Ray Peat Email Advice Depository

[Mito]

Q: What do you think is the safest approach to raising DHEA levels in a male that has below reference range DHEA according to testing?

A: Sometimes supplementing pregnenolone can do it, but 5 mg of DHEA is safe, if your thyroid, vitamin D, and other tests are normal.

 

[John Frusciante]

Still on Airplanes/dirigibles/transportation conversation:

"A couple quick questions: when you mention good rail transportation, do you mean super fast trains like the MagLev (goes up to 600km/hour in the latest tests) that are being projected by Germany and Japan?

About dirigibles: I can only read their use been declining in the past decades. Do you mean using them as a mean of transportation? In the public opinion, airplanes aren't problematic, and are preferred over dirigibles. Am I missing something?"

"Public opinion is formed to optimize profits. Even hydrogen-filled dirigibles are safe and cheap, and rapid change of time zones is seriously stressful. In 1937, the Germans were leading in commercial aviation, and the US government was subsidizing domestic airlines to carry mail, permitting them to also carry a few paying passengers. The Hindenburg disaster was essential for the development of a US owned air passenger industry. People had been reluctant to fly for safety reasons, as well as cost. The dirigibles took just over two days to cross, and provided beds and high quality dining, for $400. The first US transatlantic airline cost $375, people had to sit up for more than 24 hours, and only a few passengers were carried along with the mail. The efficiency of airlines is analogous to the efficiency of nuclear power, i.e., it rests on government subsidies. A few days ago, the Amtrack accident was the result of trying to save ten minutes on the trip from Seattle to Portland. When government money is involved, people go crazy."

 

[Ania]

The question is below, first is the answer from dr Peat, sorry for that

Have you checked your temperature and pulse rate cycles, to see if the progesterone affects them? What about your thyroid function? Low thyroid function commonly causes beast swelling. Under the influence of progesterone, estrogen is converted to the water soluble sulfate and glucuronide forms, to be excreted; if there is inflammation, from an infection or injury, these inactivated forms of estrogen can be re-activated in the inflamed tissue.


Dear dr Peat,

Whenever I take progest-e or even pregnenolone my breasts become a little swallen, tender and the right breast get painful with some sort of protrusion which I can feel with my fingers. Do you think it might be estrogen pushed into the bloodstream? Should I ignore this symptom and continue administration until it vanishes?
It happened even when I applied small amounts of progest-e topically.
I'll be grateful for any tips.

 

[cjm]

Regarding DIY ophthalmic preparations:

ME: Hi Ray,

I'm not sure if you will get this email but I couldn't find our correspondence from times I have emailed you in the past. I have a quick question for you. I am attempting to prepare an ophthalmic solution with coconut oil and liquid T3, and I want to dilute the coconut oil so the T3 can disperse fully in the solution. (The T3 I'm using isn't optimal as it contains some portion of ethanol but it is my only option and I have already experienced great relief of my symptoms (the inspiration to make this solution came from one of your articles in which you mention a T3-containing eye drop likely being more effective at restoring metabolism than oral administration of thyroid)). Do you know of a safe emulsifier I can use in my preparation? My first thought is egg yolk but I'm really out in left field here as far as safety of ophthalmic solutions is concerned.

There is a pilot study with rabbits on the efficacy of virgin coconut oil as an eye re-wetting agent, but they apparently did not alter it in any way.

https://www.hindawi.com/journals/ecam/2015/135987/

Thanks for your consideration,
Chris

RAY: I know a couple of people who have used refined coconut oil in their eyes, as a solvent for other things, and they say it isn’t especially uncomfortable, but for some people, any oil in the eye can be painful. I think an emulsifier, even egg yolk, would have an effect similar to soap in the eye.

ME: Hi again Ray, do you think tocopherols would be an eye irritant? I am curious to try small doses of fat soluble vitamins through the eye.

RAY: I’ve accidentally got some in my eye when I was putting it on for sunburn, and it was very uncomfortable.

 

[raypeatclips]

Peat not as anti fiber/anti vegetable as people make out?

Q. What would you recommend to prevent against bowel cancer?

I think fibrous foods and milk are the most protective things, along with good thyroid function, and avoiding polyunsaturated fats.

Q. I have seen you speak about "undercooked" vegetables as a problem, and many studies you reference regarding fiber are concerned with things such as oat bran, not vegetables. I wondered how bad you thought "normal" vegetables such as peppers, courgettes, tomatoes etc were, if they were cooked to excess, for example 30+ minutes to be eaten as a side alongside a meal?

They are very good nutritionally, if they don’t cause any problems of intestinal inflammation (even cooked peppers and tomatoes cause problems for lots of people).

Q. Are you aware of any other vegetables with a reputation for causing inflammation?

Green leaves used in salads very commonly support bacterial overgrowth; seeds, nuts, and grains, and some starchy vegetables, especially when they aren’t cooked until they soften.

 

[raypeatclips]

Q.
I have seen you mention spermidine is a polyamine. There are studies regarding spermidine as being positive such as

Spermidine Prolongs Lifespan and Prevents Liver Fibrosis and Hepatocellular Carcinoma by Activating MAP1S-Mediated Autophagy

Spermidine Prolongs Lifespan and Prevents Liver Fibrosis and Hepatocellular Carcinoma by Activating MAP1S-Mediated Autophagy

"Extending recent evidence that orally administered spermidine can extend lifespan in mice, we determined that life extension of up to 25% can be produced by lifelong administration, which also reduced liver fibrosis and HCC foci as induced by chemical insults."

I also was shown a piece looking at the polyamine content of various foods, and found mushrooms particularly high
http://foodandnutrition.jacobspublishers.com/images/FoodNutrition/J_J_Food_Nutri_4_1_029.pdf

Yet mushrooms seem anti cancer, and good for various factors of health. Is spermidine a cause for concern in the diet? What do you make of all this?

I think the total effect of each food has to be considered. Polyamines are needed for renewal and regeneration, but I don’t think it’s safe to supplement them.

1. Med Sci (Basel). 2017 Dec 7;5(4). pii: E32. doi: 10.3390/medsci5040032.
Investigation of Polyamine Metabolism and Homeostasis in Pancreatic Cancers.
Massaro C(1), Thomas J(2), Phanstiel Iv O(3).
(1)Department of Medical Education, College of Medicine, University of Central
Florida, Orlando, FL 32826-3227, USA. [email protected].
(2)Department of Medical Education, College of Medicine, University of Central
Florida, Orlando, FL 32826-3227, USA. [email protected].
(3)Department of Medical Education, College of Medicine, University of Central
Florida, Orlando, FL 32826-3227, USA. [email protected].

Pancreatic cancers are currently the fourth leading cause of cancer-related death
and new therapies are desperately needed. The most common pancreatic cancer is
pancreatic ductal adenocarcinoma (PDAC). This report describes the development of
therapies, which effectively deplete PDAC cells of their required polyamine
growth factors. Of all human tissues, the pancreas has the highest level of the
native polyamine spermidine. To sustain their high growth rates, PDACs have
altered polyamine metabolism, which is reflected in their high intracellular
polyamine levels and their upregulated import of exogenous polyamines. To
understand how these cancers respond to interventions that target their specific
polyamine pools, L3.6pl human pancreatic cancer cells were challenged with
specific inhibitors of polyamine biosynthesis. We found that pancreatic cell
lines have excess polyamine pools, which they rebalance to address deficiencies
induced by inhibitors of specific steps in polyamine biosynthesis (e.g.,
ornithine decarboxylase (ODC), spermidine synthase (SRM), and spermine synthase
(SMS)). We also discovered that combination therapies targeting ODC, SMS, and
polyamine import were the most effective in reducing intracellular polyamine
pools and reducing PDAC cell growth. A combination therapy containing
difluoromethylornithine (DFMO, an ODC inhibitor) and a polyamine transport
inhibitor (PTI) were shown to significantly deplete intracellular polyamine
pools. The additional presence of an SMS inhibitor as low as 100 nM was
sufficient to further potentiate the DFMO + PTI treatment.


2. Mini Rev Med Chem. 2017 Sep 27. doi: 10.2174/1389557517666170927130526. [Epub
ahead of print]
Ornithine Decarboxylase Inhibition: A strategy to combat various diseases.
Rai PR(1), Somani RR(1), Kandpile PS(1).
(1)Vivekanand Education Society`s College Of Pharmacy - Department of
Pharmaceutical Chemistry Mumbai, Maharashtra. India.

Ornithine decarboxylase is the first enzyme in the polyamine biosynthetic
pathway. It is the rate-limiting enzyme which is included in the change of
ornithine to putrescine which is the first polyamine. Polyamines (putrescine,
spermidine, spermine) are natural and synthetic compounds which contains two or
more amino group. Polyamines are highly implicated in cellular functions such as
cell-growth & multiplication, DNA stabilization, gene transcription and
translation, ion-channel activity, etc. Elevated levels of polyamines were found
in highly proliferating tumour cells. Hence inhibition of this enzyme was found
useful in cancer. α-DL-difluoromethylornithine(DFMO) (Eflornithine) an
enzyme-activated irreversible inhibitor was the first of this type. However its
use as an anticancer agent did not continue for long due to various reasons.
Polyamines were also found to play important role in other infectious
microorganism. Eflornithine is successfully used in diseases such as African
sleeping sickness and are being researched against number of tropical diseases.
It is widely used against hirsutism in women. Various other product (putrscine)
based analogues and transition state or PLP (cofactor) based analagoues are being
synthesized against diseases such as Leishmaniasis, malaria and others discussed
in the article.



3. Biochem Biophys Res Commun. 2017 Aug 26;490(3):861-867. doi:
10.1016/j.bbrc.2017.06.132. Epub 2017 Jun 22.
Helicobacter pylori does not use spermidine synthase to produce spermidine.
Zhang H(1), Au SWN(2).
(1)Centre for Protein Science and Crystallography, School of Life Sciences, The
Chinese University of Hong Kong, Hong Kong.
(2)Centre for Protein Science and Crystallography, School of Life Sciences, The
Chinese University of Hong Kong, Hong Kong. Electronic address:
[email protected].

Helicobacter pylori is the primary pathogen associated to gastritis and gastric
cancer. Growth of H. pylori depends on the availability of spermidine in vivo.
Interestingly, the genome of H. pylori contains an incomplete set of genes for
the classical pathway of spermidine biosynthesis. It is thus not clear whether
some other genes remained in the pathway would have any functions in spermidine
biosynthesis. Here, we study spermidine synthase, which is responsible for the
final catalytic process in the classical route. Protein sequence alignment
reveals that H. pylori SpeE (HpSpeE) lacks key residues for substrate binding. By
using isothermal titration calorimetry, we show that purified recombinant HpSpeE
does not interact with the putative substrates putrescine and decarboxylated
S-adenosylmethionine, and the product spermidine. High performance liquid
chromatography analysis further demonstrates that HpSpeE has no detectable
in vitro enzymatic activity. Additionally, intracellular spermidine level in
speE-null mutant strain is comparable to that in the wild type strain.
Collectively, our results suggest that HpSpeE is functionally distinct from
spermidine production. H. pylori may instead employ the alternative pathway for
spermidine synthesis which is dominantly exploited by other human gut microbes.



4. Anticancer Drugs. 2017 Apr;28(4):457-464. doi: 10.1097/CAD.0000000000000465.
Extracellular polyamines-induced proliferation and migration of cancer cells by
ODC, SSAT, and Akt1-mediated pathway.
Dai F(1), Yu W, Song J, Li Q, Wang C, Xie S.
(1)aInstitute of Chemical Biology, College of Pharmacy bThe Key Laboratory of
Natural Medicine and Immuno-Engineering, Henan University, Kaifeng, China.

High levels of polyamines were observed and were related to a poor prognosis in
cancer patients. However, the mechanism is not obvious. The aim of this study is
to mimic the extracellular polyamines in a tumor microenviroment and to explore
the role of extracellular polyamines in the proliferation and migration of cancer
cells. Three different concentrations of polyamines composed of putrescine,
spermidine, and spermine were used. Colony formation assay, wound healing assay,
and transwell migration assay were performed. Akt1-overexpression cells were
constructed. The related protein expression was examined using a western blot. In
this study, polyamines promoted colony formation and cell migration in a
concentration-dependent and time-dependent manner. Polyamines upregulated the
expression of ornithine decarboxylase (ODC), SSAT, Akt1, Akt, hypoxia-inducible
factors-1α, vascular endothelial growth factor, and matrix metalloproteinases,
and downregulated p27 expression. The effects of combination of polyamines and
Akt1 overexpression on colony formation and migration were more obvious than the
effects of Akt1 overexpression alone. In Akt1-overexpression cells, polyamines
also upregulated the expression of ODC, SSAT, hypoxia-inducible factors-1α,
vascular endothelial growth factor, and matrix metalloproteinases and
downregulated p27 expression. In conclusion, extracellular polyamines induced
proliferation and cancer cell migration by inducing ODC and SSAT expression, and
the Akt1-mediated pathway.


5. Oncotarget. 2017 Jan 3;8(1):1092-1109. doi: 10.18632/oncotarget.13582.
Spermidine/spermine N1-acetyltransferase regulates cell growth and metastasis via
AKT/β-catenin signaling pathways in hepatocellular and colorectal carcinoma
cells.
Wang C(1), Ruan P(1), Zhao Y(1), Li X(1), Wang J(1), Wu X(1), Liu T(1), Wang
S(1), Hou J(1), Li W(1), Li Q(2), Li J(2), Dai F(2), Fang D(1), Wang C(2), Xie
S(1).
(1)Institute of Chemical Biology, College of Pharmacy, Henan University, Kaifeng,
475004, China.
(2)The Key Laboratory of Natural Medicine and Immuno-Engineering, Henan
University, Kaifeng, 475004, China.

Hepatocellular carcinoma (HCC) and colorectal cancer (CRC) are among the most
common cancers across the world. Therefore, identifying the potential molecular
mechanisms that promote HCC and CRC progression and metastasis are urgently
needed. Spermidine/spermine N1-acetyltransferase (SSAT) is a catabolic enzyme
that acetylates the high-order polyamines spermine and spermidine, thus
decreasing the cellular content of polyamines. Several publications have
suggested that depletion of intracellular polyamines inhibited tumor progression
and metastasis in various cancer cells. However, whether and how SSAT regulates
cell growth, migration and invasion in hepatocellular and colorectal carcinoma
cells remains unclear. In this study, depletion of polyamines mediated by SSAT
not only attenuated the tumor cell proliferation but also dramatically inhibited
cell migration and invasion in hepatocellular and colorectal carcinoma cells.
Subsequent investigations revealed introduction of SSAT into HepG2, SMMC7721
hepatocellular carcinoma cells and HCT116 colorectal carcinoma cells
significantly suppressed p-AKT, p-GSK3β expression as well as β-catenin nuclear
translocation, while inhibition of GSK3β activity or exogenous polyamines could
restore SSAT-induced decreases in the protein expression of p-AKT, p-GSK3β and
β-catenin. Conversely, knockdown of SSAT in Bel7402 hepatocellular carcinoma
cells and HT-29 colorectal carcinoma cells which expressed high levels of SSAT
endogenously significantly promoted the expression of p-AKT, p-GSK3β as well as
β-catenin nuclear translocation. Taken together, our results indicated depletion
of polyamines by SSAT significantly inhibited cell proliferation, migration and
invasion through AKT/GSK3β/β-catenin signaling pathway in hepatocellular
carcinoma and colorectal cancer cells.



6. Proc Natl Acad Sci U S A. 2016 Nov 1;113(44):E6806-E6812. Epub 2016 Oct 3.

Activation of SAT1 engages polyamine metabolism with p53-mediated ferroptotic
responses.

Ou Y(1)(2), Wang SJ(1)(2), Li D(1)(2), Chu B(1)(2), Gu W(3)(2).

Author information:
(1)Institute for Cancer Genetics, Department of Pathology and Cell Biology,
College of Physicians and Surgeons, Columbia University, New York, NY 10032.
(2)Herbert Irving Comprehensive Cancer Center, College of Physicians and
Surgeons, Columbia University, New York, NY 10032.
(3)Institute for Cancer Genetics, Department of Pathology and Cell Biology,
College of Physicians and Surgeons, Columbia University, New York, NY 10032;
[email protected].

Although p53-mediated cell-cycle arrest, senescence, and apoptosis remain
critical barriers to cancer development, the emerging role of p53 in cell
metabolism, oxidative responses, and ferroptotic cell death has been a topic of
great interest. Nevertheless, it is unclear how p53 orchestrates its activities
in multiple metabolic pathways into tumor suppressive effects. Here, we
identified the SAT1 (spermidine/spermine N1-acetyltransferase 1) gene as a
transcription target of p53. SAT1 is a rate-limiting enzyme in polyamine
catabolism critically involved in the conversion of spermidine and spermine back
to putrescine. Surprisingly, we found that activation of SAT1 expression induces
lipid peroxidation and sensitizes cells to undergo ferroptosis upon reactive
oxygen species (ROS)-induced stress, which also leads to suppression of tumor
growth in xenograft tumor models. Notably, SAT1 expression is down-regulated in
human tumors, and CRISPR-cas9-mediated knockout of SAT1 expression partially
abrogates p53-mediated ferroptosis. Moreover, SAT1 induction is correlated with
the expression levels of arachidonate 15-lipoxygenase (ALOX15), and SAT1-induced
ferroptosis is significantly abrogated in the presence of PD146176, a specific
inhibitor of ALOX15. Thus, our findings uncover a metabolic target of p53
involved in ferroptotic cell death and provide insight into the regulation of
polyamine metabolism and ferroptosis-mediated tumor suppression.



7. Autophagy. 2016 Oct 2;12(10):1962-1964. Epub 2016 Aug 17.

Autophagy induction for the treatment of cancer.

Pietrocola F(1)(2)(3)(4), Pol J(1)(2)(3)(4), Vacchelli E(1)(2)(3)(4)(5), Baracco
EE(1)(2)(3)(4)(5)(6), Levesque S(1)(2)(3)(4)(5)(6), Castoldi
F(1)(2)(3)(4)(5)(6)(7), Maiuri MC(1)(2)(3)(4)(5), Madeo F(8)(9), Kroemer
G(1)(2)(3)(4)(6)(10)(11)(12).

Author information:
(1)a Gustave Roussy Cancer Campus , Villejuif , France.
(2)b INSERM , U1138 , Paris , France.
(3)c Equipe 11 labellisée par la Ligue Nationale contre le Cancer , Centre de
Recherche des Cordeliers , Paris , France.
(4)d Université Paris Descartes/Paris V , Sorbonne Paris Cité, Paris , France.
(5)e Université Pierre & Marie Curie , Paris , France.
(6)f Université Paris-Sud/Paris XI , Faculté de Médecine , Kremlin-Bicêtre ,
France.
(7)g Sotio a.c. , Prague , Czech Republic.
(8)h BioTechMed-Graz , Graz , Austria.
(9)i BioTechMed-Graz Graz , Austria.
(10)j Metabolomics and Cell Biology Platforms , Gustave Roussy Cancer Campus ,
Villejuif , France.
(11)k Pôle de Biologie , Hôpital Européen Georges Pompidou, AP-HP , Paris ,
France.
(12)l Department of Women's and Children's Health , Karolinska University
Hospital , Stockholm , Sweden.

Cancer can be viewed in 2 rather distinct ways, namely (i) as a cell-autonomous
disease in which malignant cells have escaped control from cell-intrinsic
barriers against proliferation and dissemination or (ii) as a systemic disease
that involves failing immune control of aberrant cells. Since
macroautophagy/autophagy generally increases the fitness of cells as well as
their resistance against endogenous or iatrogenic (i.e., relating to illness due
to medical intervention) stress, it has been widely proposed that inhibition of
autophagy would constitute a valid strategy for sensitizing cancer cells to
chemotherapy or radiotherapy. Colliding with this cell-autonomous vision,
however, we found that immunosurveillance against transplantable,
carcinogen-induced or genetically engineered cancers can be improved by
pharmacologically inducing autophagy with caloric restriction mimetics. This
positive effect depends on autophagy induction in cancer cells and is mediated by
alterations in extracellular ATP metabolism, namely increased release of
immunostimulatory ATP and reduced adenosine-dependent recruitment of
immunosuppressive regulatory T cells into the tumor bed. The combination of
autophagy inducers and chemotherapeutic agents is particularly efficient in
reducing cancer growth through the stimulation of CD8+ T lymphocyte-dependent
anticancer immune responses.


8. Cancer Cell. 2016 Jul 11;30(1):147-160. doi: 10.1016/j.ccell.2016.05.016.
Caloric Restriction Mimetics Enhance Anticancer Immunosurveillance.
Pietrocola F(1), Pol J(2), Vacchelli E(1), Rao S(3), Enot DP(4), Baracco EE(5),
Levesque S(5), Castoldi F(6), Jacquelot N(7), Yamazaki T(7), Senovilla L(2),
Marino G(1), Aranda F(1), Durand S(4), Sica V(5), Chery A(4), Lachkar S(5), Sigl
V(3), Bloy N(5), Buque A(5), Falzoni S(8), Ryffel B(9), Apetoh L(10), Di Virgilio
F(8), Madeo F(11), Maiuri MC(5), Zitvogel L(7), Levine B(12), Penninger JM(3),
Kroemer G(13).
(1)Gustave Roussy Cancer Campus, 94800 Villejuif, France; INSERM, U1138, 75006
Paris, France; Equipe 11 labellisée par la Ligue Nationale contre le Cancer,
Centre de Recherche des Cordeliers, 75006 Paris, France.
(2)Gustave Roussy Cancer Campus, 94800 Villejuif, France; INSERM, U1138, 75006
Paris, France; Equipe 11 labellisée par la Ligue Nationale contre le Cancer,
Centre de Recherche des Cordeliers, 75006 Paris, France; Université Paris
Descartes/Paris V, Sorbonne Paris Cité, 75006 Paris, France.
(3)Institute for Molecular Biotechnology of the Austrian Academy of Sciences,
1030 Vienna, Austria.
(4)Gustave Roussy Cancer Campus, 94800 Villejuif, France; Metabolomics and Cell
Biology Platforms, Gustave Roussy Cancer Campus, 94800 Villejuif, France.
(5)Gustave Roussy Cancer Campus, 94800 Villejuif, France; INSERM, U1138, 75006
Paris, France; Equipe 11 labellisée par la Ligue Nationale contre le Cancer,
Centre de Recherche des Cordeliers, 75006 Paris, France; Université
Paris-Sud/Paris-Saclay, Faculté de Médecine, 94276 Kremlin-Bicêtre, France.
(6)Gustave Roussy Cancer Campus, 94800 Villejuif, France; INSERM, U1138, 75006
Paris, France; Equipe 11 labellisée par la Ligue Nationale contre le Cancer,
Centre de Recherche des Cordeliers, 75006 Paris, France; Université
Paris-Sud/Paris-Saclay, Faculté de Médecine, 94276 Kremlin-Bicêtre, France; Sotio
a.c., 17000 Prague, Czech Republic.
(7)Gustave Roussy Cancer Campus, 94800 Villejuif, France; Université
Paris-Sud/Paris-Saclay, Faculté de Médecine, 94276 Kremlin-Bicêtre, France;
INSERM U1015, Gustave Roussy Cancer Campus, 94800 Villejuif, France; Center of
Clinical Investigations in Biotherapies of Cancer (CICBT) 1428, 94800 Villejuif,
France.
(8)Department of Morphology, Surgery and Experimental Medicine University of
Ferrara, 44121 Ferrara, Italy.
(9)UMR7355, CNRS and University, 45067 Orléans, France; Institute of Infectious
Disease and Molecular Medicine, University of Cape Town, 7925 Cape Town, South
Africa.
(10)INSERM, U866, 21078 Dijon, France; Faculté de Médecine, Université de
Bourgogne, 21078 Dijon, France; Centre Georges François Leclerc, 21000 Dijon,
France.
(11)Institute of Molecular Biosciences, NAWI Graz, University of Graz,
Humboldtstraße 50, 8010 Graz, Austria; BioTechMed-Graz, Humboldtstraße 50, 8010
Graz, Austria.
(12)Department of Internal Medicine, Center for Autophagy Research, Howard Hughes
Medical Institute, University of Texas Southwestern Medical Center, Dallas, TX
73590, USA.
(13)Gustave Roussy Cancer Campus, 94800 Villejuif, France; INSERM, U1138, 75006
Paris, France; Equipe 11 labellisée par la Ligue Nationale contre le Cancer,
Centre de Recherche des Cordeliers, 75006 Paris, France; Université Paris
Descartes/Paris V, Sorbonne Paris Cité, 75006 Paris, France; Metabolomics and
Cell Biology Platforms, Gustave Roussy Cancer Campus, 94800 Villejuif, France;
Pôle de Biologie, Hôpital Européen Georges Pompidou, AP-HP, 75015 Paris, France;
Department of Women's and Children's Health, Karolinska Institute, Karolinska
University Hospital, Stockholm, Sweden; INSERM U1138, Centre de Recherche des
Cordeliers, 75006 Paris, France. Electronic address: [email protected].
Comment in
Cancer Cell. 2016 Jul 11;30(1):13-5.
Caloric restriction mimetics (CRMs) mimic the biochemical effects of nutrient
deprivation by reducing lysine acetylation of cellular proteins, thus triggering
autophagy. Treatment with the CRM hydroxycitrate, an inhibitor of ATP citrate
lyase, induced the depletion of regulatory T cells (which dampen anticancer
immunity) from autophagy-competent, but not autophagy-deficient, mutant
KRAS-induced lung cancers in mice, thereby improving anticancer
immunosurveillance and reducing tumor mass. Short-term fasting or treatment with
several chemically unrelated autophagy-inducing CRMs, including hydroxycitrate
and spermidine, improved the inhibition of tumor growth by chemotherapy in vivo.
This effect was only observed for autophagy-competent tumors, depended on the
presence of T lymphocytes, and was accompanied by the depletion of regulatory
T cells from the tumor bed.



9. Amino Acids. 2016 Oct;48(10):2401-10. doi: 10.1007/s00726-016-2256-6. Epub 2016
May 14.
Intracellular sources of ornithine for polyamine synthesis in endothelial cells.
Li H(1), Meininger CJ(2), Bazer FW(1), Wu G(3)(4).

(1)Department of Animal Science, Texas A&M University, College Station, TX,
77843, USA.
(2)Department of Medical Physiology, Texas A&M University, College Station, TX,
77843, USA.
(3)Department of Animal Science, Texas A&M University, College Station, TX,
77843, USA. [email protected].
(4)Department of Medical Physiology, Texas A&M University, College Station, TX,
77843, USA. [email protected].

Polyamines are essential for proliferation of endothelial cells (EC) and
angiogenesis. This study was conducted to identify the metabolic source(s) of
ornithine for polyamine synthesis in EC, using N(ω)-hydroxy-nor-L-arginine
(Nor-NOHA, an inhibitor of arginase) and gabaculine (an inhibitor of ornithine
aminotransferase; OAT). Nor-NOHA inhibited arginase with an IC50 value of 10 µM
for intact EC. Nor-NOHA (0.5 mM) alone inhibited arginase activity in EC by 98 %,
increased total cellular concentrations of arginine by 14 %, and decreased total
cellular concentrations of ornithine, putrescine and spermidine by 17, 65 and
74 %, respectively. Arginine and glutamine contributed to 73 and 26 % of the
ornithine produced by EC, respectively. Gabaculine (1 mM) alone decreased the
total cellular concentrations of arginine, ornithine, putrescine, and spermidine
by 14, 96, 32, and 42 %, respectively. A combination of both Nor-NOHA and
gabaculine completely blocked ornithine production in EC, resulting in no
detectable cellular ornithine and almost complete depletion of cellular
putrescine and spermidine. Addition of 0.5 mM ornithine restored the
intracellular concentrations of polyamines in EC treated with Nor-NOHA plus
gabaculine, indicating that Nor-NOHA and gabaculine did not inhibit ornithine
decarboxylase activity. Our results suggest that the arginase and OAT pathways
are the exclusive sources of ornithine in EC when there is little extracellular
ornithine and that there is intracellular compartmentalization of arginine and
ornithine for endothelial synthesis of polyamines. These novel findings may have
important implications for improving placental vascular growth, wound healing,
and cancer therapy.


10. Bioessays. 2016 Jun;38(6):498-507. doi: 10.1002/bies.201500195. Epub 2016 Apr 29.

Polyamine signal through gap junctions: A key regulator of proliferation and
gap-junction organization in mammalian tissues?

Hamon L(1), Savarin P(2), Pastré D(1).

Author information:
(1)Laboratoire Structure-Activité des Biomolécules Normales et Pathologiques,
INSERM U1204 and Université Evry-Val d'Essonne, Evry, France.
(2)Centre National de Recherche Scientifique (CNRS), Equipe Spectroscopie des
Biomolécules et des Milieux Biologiques (SBMB), Université Paris 13, Sorbonne
Paris Cité, Laboratoire Chimie, Structures, Propriétés de Biomatériaux et
d'Agents Thérapeutiques (CSPBAT), Unité Mixte de Recherche (UMR) 7244, Bobigny,
France.

We propose that interaction rules derived from polyamine exchange in connected
cells may explain the spatio-temporal organization of gap junctions observed
during tissue regeneration and tumorigenesis. We also hypothesize that polyamine
exchange can be considered as signal that allows cells to sense the proliferation
status of their neighbors. Polyamines (putrescine, spermidine, and spermine) are
indeed small aliphatic polycations that serve as fuels to sustain elevated
proliferation rates of the order observed in cancer cells. Based on recent
reports, we consider here that polyamines can be exchanged through gap junction
channels between mammalian cells. Such intercellular exchange of polyamines has
critical consequences on the local control of growth. In line with this
hypothesis, the complex protein network that keeps polyamine levels finely tuned
in mammalian cells can translate polyamine efflux or influx into integrated
signals controlling transcription, translation, and cell communications.



11. Cancer Lett. 2016 Jul 10;377(1):44-54. doi: 10.1016/j.canlet.2016.04.027. Epub
2016 Apr 20.

Calcium sensing receptor suppresses human pancreatic tumorigenesis through a
novel NCX1/Ca(2+)/β-catenin signaling pathway.

Tang B(1), Chow JY(2), Dong TX(2), Yang SM(1), Lu DS(3), Carethers JM(4), Dong
H(5).

Author information:
(1)Department of Gastroenterology, Xinqiao Hospital, Third Military Medical
University, Chongqing, China.
(2)Department of Medicine, University of California, San Diego, CA, USA.
(3)Cancer Research Center, Shenzhen University, Shenzhen, China.
(4)Department of Internal Medicine, University of Michigan, Ann Arbor, MI, USA.
Electronic address: [email protected].
(5)Department of Gastroenterology, Xinqiao Hospital, Third Military Medical
University, Chongqing, China; Department of Medicine, University of California,
San Diego, CA, USA. Electronic address: [email protected].

The calcium sensing receptor (CaSR) is functionally expressed in normal human
pancreases, but its pathological role in pancreatic tumorigenesis is currently
unknown. We sought to investigate the role of CaSR in pancreatic cancer (PC) and
the underlying molecular mechanisms. We revealed that the expression of CaSR was
consistently downregulated in the primary cancer tissues from PC patients, which
was correlated with tumor size, differentiation and poor survival of the
patients. CaSR activation markedly suppressed pancreatic tumorigenesis in vitro
and in vivo likely through the Ca(2+) entry mode of Na(+)/Ca(2+) exchanger 1
(NCX1) to induce Ca(2+) entry into PC cells. Moreover, NCX1-mediated Ca(2+) entry
resulted in Ca(2+)-dependent inhibition of β-catenin signaling in PC cells,
eventually leading to the inhibition of pancreatic tumorigenesis. Collectively,
we demonstrate for the first time that CaSR exerts a suppressive function in
pancreatic tumorigenesis through a novel NCX1/Ca(2+)/β-catenin signaling pathway.
Targeting this specific signaling pathway could be a potential therapeutic
strategy for PC.




12. Biochem J. 2016 May 15;473(10):1433-41. doi: 10.1042/BCJ20160134. Epub 2016 Mar
21.

Triethylenetetramine modulates polyamine and energy metabolism and inhibits
cancer cell proliferation.

Hyvönen MT(1), Ucal S(1), Pasanen M(1), Peräniemi S(1), Weisell J(1), Khomutov
M(2), Khomutov AR(2), Vepsäläinen J(1), Alhonen L(1), Keinänen TA(3).

Author information:
(1)School of Pharmacy, University of Eastern Finland, Yliopistonranta 1C, 70210
Kuopio, Finland.
(2)Engelhardt Institute of Molecular Biology, Russian Academy of Sciences,
Vavilov St 32, 119991 Moscow, Russia.
(3)School of Pharmacy, University of Eastern Finland, Yliopistonranta 1C, 70210
Kuopio, Finland [email protected].

Polyamine metabolism is an attractive anticancer drug target, since polyamines
are absolutely required for cellular proliferation, and increased levels of
polyamines and their biosynthetic enzyme ornithine decarboxylase (ODC) are
associated with cancer. Triethylenetetramine (TETA) is a charge-deficient
isosteric analogue of the polyamine spermidine (Spd) and a Cu(II)-chelating
compound used for the treatment of Wilson's disease, and it has been implicated
as a potential anticancer therapeutic drug. In the present study, we studied the
effects of TETA in comparison with two other Cu(II)-chelators, D-penicillamine
(PA) and tetrathiomolybdate (TTM), on polyamine metabolism in DU145 prostate
carcinoma, MCF-7 breast carcinoma and JEG-3 choriocarcinoma cells. TETA induced
antizyme, down-regulated ODC and inhibited [(14)C] Spd uptake. Moreover, it
completely prevented α-difluoromethylornithine (DFMO)-induced increase in [(14)C]
Spd uptake, and inhibited [(14)C] putrescine (Put) uptake and ODC activity
in vivo Seven-day treatment of DU145 cells with TETA caused growth cessation by
reducing intracellular polyamine levels and suppressing the formation of
hypusinated eukaryotic translation initiation factor 5A (eIF5A). TETA or its
N-acetylated metabolites also inhibited spermine (Spm), diamine and
semicarbazide-sensitive amine oxidases and decreased the level of intracellular
reactive oxygen species. Moreover, TETA inhibited the utilization of Put as
energy source via the tricarboxylic acid (TCA) cycle, as indicated by decreased
production of (14)CO2 from [(14)C] Put. These results indicate that TETA attacks
multiple proven anticancer drug targets not attributed to copper chelation, which
warrants further studies to reveal its potential in cancer chemoprevention and
cure.




13. Bioorg Med Chem Lett. 2016 Mar 15;26(6):1629-1632. doi:
10.1016/j.bmcl.2016.01.083. Epub 2016 Feb 4.

Synthesis and anticancer evaluation of spermatinamine analogues.

Moosa BA(1), Sagar S(2), Li S(3), Esau L(2), Kaur M(4), Khashab NM(3).

Author information:
(1)Controlled Release and Delivery (CRD) Lab, Chemical Life Sciences and
Engineering, King Abdullah University of Science and Technology, Thuwal
23955-6900, Saudi Arabia; Center for Advanced Membranes and Porous Materials,
King Abdullah University of Science and Technology, Thuwal 23955-6900, Saudi
Arabia. Electronic address: [email protected].
(2)Biomolecular Lab, Computational Bioscience Research Center, King Abdullah
University of Science and Technology, Thuwal 23955-6900, Saudi Arabia.
(3)Controlled Release and Delivery (CRD) Lab, Chemical Life Sciences and
Engineering, King Abdullah University of Science and Technology, Thuwal
23955-6900, Saudi Arabia; Center for Advanced Membranes and Porous Materials,
King Abdullah University of Science and Technology, Thuwal 23955-6900, Saudi
Arabia.
(4)Biomolecular Lab, Computational Bioscience Research Center, King Abdullah
University of Science and Technology, Thuwal 23955-6900, Saudi Arabia; School of
Molecular and Cell Biology, University of the Witwatersrand, Private Bag 3, Wits,
2050, Johannesburg, South Africa.

Spermatinamine was isolated from an Australian marine sponge, Pseudoceratina sp.
as an inhibitor of isoprenylcysteine carboxyl methyltransferase (Icmt), an
attractive and novel anticancer target. Herein, we report the synthesis of
spermatinamine analogues and their cytotoxic evaluation against three human
cancer cell lines, that is, cervix adenocarcinoma (HeLa), breast adenocarcinoma
(MCF-7), and prostate carcinoma (DU145). Analogues 12, 14 and 15 were found to be
the most potent against one or more cell lines with the IC50 values in the range
of 5-10 μM. The obtained results suggested that longer polyamine linker along
with aromatic oxime substitution provided the most potent analogue compounds
against cancer cell lines.



14. Bioorg Med Chem Lett. 2016 Feb 15;26(4):1145-50. doi: 10.1016/j.bmcl.2016.01.048.
Epub 2016 Jan 19.

Synthesis of minoxidil conjugates and their evaluation as HL-60 differentiation
agents.

Stoica S(1), Magoulas GE(2), Antoniou AI(2), Suleiman S(1), Cassar A(1), Gatt
L(1), Papaioannou D(2), Athanassopoulos CM(3), Schembri-Wismayer P(4).

Author information:
(1)Anatomy Department, Faculty of Medicine and Surgery, University of Malta,
Malta.
(2)Laboratory of Synthetic Organic Chemistry, Department of Chemistry, University
of Patras, GR-26504 Patras, Greece.
(3)Laboratory of Synthetic Organic Chemistry, Department of Chemistry, University
of Patras, GR-26504 Patras, Greece. Electronic address:
[email protected].
(4)Anatomy Department, Faculty of Medicine and Surgery, University of Malta,
Malta. Electronic address: [email protected].

Activation of minoxidil (MNX) with N,N'-carbonyldiimidazole and coupling with
natural polyamines (PAs) and commercially available aliphatic or aromatic amines
provided a series of new conjugates which were evaluated for their ability to
induce differentiation to HL-60 acute myeloid leukemia cancer cells, using a
modified NBTZ reduction test. Although neither MNX nor 4,4'-methylenedianiline
(MDA) or 2,7-diaminofluorene (DAF), alone or in combination, had any effect, the
MNX-spermine (SPM) conjugate (11) and the conjugates 7 and 8 of MNX with MDA and
DAF exhibited a differentiation-inducing effect at a concentration of 10 μM
without being toxic on proliferating human peripheral blood mononuclear cells.




15. Amino Acids. 2016 Apr;48(4):1003-12. doi: 10.1007/s00726-015-2143-6. Epub 2015
Dec 24.

Decreased sensitivity to aspirin is associated with altered polyamine metabolism
in human prostate cancer cells.

Li J(1)(2), Cameron GA(1), Wallace HM(3).

Author information:
(1)Division of Applied Medicine, School of Medicine and Dentistry and School of
Medical Sciences, University of Aberdeen, Aberdeen, UK.
(2)Cancer Research Centre, Western General Hospital, University of Edinburgh,
Edinburgh, UK.
(3)Division of Applied Medicine, School of Medicine and Dentistry and School of
Medical Sciences, University of Aberdeen, Aberdeen, UK. [email protected].

Aspirin is a well-known analgesic, anti-inflammatory and antipyretic drug and is
recognised as a chemopreventative agent in cardiovascular disease and, more
recently, in colorectal cancer. Although several studies indicate that aspirin is
capable of reducing the risk of developing cancers, there is a lack of convincing
evidence that aspirin can prevent prostate cancer in man. In this study, aspirin
was shown to be an effective inhibitor of the growth of human prostate cancer
cells. In order to investigate the link between polyamine catabolism and the
effects of aspirin we used a "Tet off" system that induced the activity of
spermidine/spermine N (1)-acetyltransferase (SSAT) in human prostate cancer cells
(LNCap). Treatment with aspirin was found to decrease induced SSAT activity in
these cells. A negative correlation was observed between increased polyamine
catabolism via increased SSAT activity and the sensitivity to aspirin. In the
presence of increased SSAT activity high amounts of N (1)-acetylspermidine and
putrescine were observed. These cells were also found to grow more slowly than
the non-induced cells. The results indicate that SSAT and its related polyamine
metabolism may play a key role in sensitivity of cancer cells to aspirin and
possibly other NSAIDs and this may have implications for the development of novel
chemopreventative agents.




16. Tumour Biol. 2016 Jan;37(1):1159-71. doi: 10.1007/s13277-015-3912-2. Epub 2015
Aug 16.

Targeting polyamine biosynthetic pathway through RNAi causes the abrogation of
MCF 7 breast cancer cell line.

Gupta ED(1), Pachauri M(2), Ghosh PC(2), Rajam MV(3).

Author information:
(1)Department of Genetics, University of Delhi South Campus, Benito Juarez Road,
New Delhi, 110021, India.
(2)Department of Biochemistry, University of Delhi South Campus, Benito Juarez
Road, New Delhi, 110021, India.
(3)Department of Genetics, University of Delhi South Campus, Benito Juarez Road,
New Delhi, 110021, India. [email protected].

The diamine putrescine and polyamines, spermidine (triamine) and spermine
(tetraamine) are small organic polycations that play an indispensable role in key
cellular processes such as the regulation of growth, differentiation, and
macromolecular functions. Elevated levels of polyamines (PAs) have been shown to
be one of the major factors involved in carcinogenesis. In this study, specific
silencing of the expression of three genes of PA biosynthesis pathway, ornithine
decarboxylase (ODC), S-adenosylmethionine decarboxylase (SAMDC), and spermidine
synthase (SPDSYN) was achieved using RNA interference in MCF 7 breast cancer cell
line. For optimizing the effective small interfering nucleic acid (siNA), three
variants of ODC siNA [siRNA, locked nucleic acid (LNA)-modified siRNA, and
siHybrid (RNA and DNA hybrid)] were used and a dose- and time-dependent study was
conducted. The PA biosynthetic genes were targeted individually and in
combination. RNAi-mediated reduction in the expression of PA biosynthesis genes
resulted in distorted cell morphology, reduced cancer cell viability, and
migration characteristic. The most promising results were observed with the
combined treatment of siSPDSYN and siODC with 83 % cell growth inhibition. On
analyzing the messenger RNA (mRNA) expression profile of the cell cycle and
apoptosis-related genes, it was observed that RNAi against PA biosynthetic genes
downregulated the expression of CDK8, CCNE2, CCNH, CCNT1, CCNT2, CCNF, PCNA,
CCND1, and CDK2, and upregulated the expression of E2F4, BAX, FAS, TP53, CDKN1A,
BAK1, CDKN1B, ATM, GRANB, and ATR genes when compared with control-transfected
cells. These results suggest that the targeting polyamine biosynthesis through
RNAi approach could be a promising strategy for breast cancer therapy and might
be extended for therapy of other cancers.



17. J Mol Biol. 2015 Oct 23;427(21):3389-406. doi: 10.1016/j.jmb.2015.06.020. Epub
2015 Jul 5.

Remaining Mysteries of Molecular Biology: The Role of Polyamines in the Cell.

Miller-Fleming L(1), Olin-Sandoval V(1), Campbell K(1), Ralser M(2).

Author information:
(1)Department of Biochemistry and Cambridge Systems Biology Centre, University of
Cambridge, Cambridge CB2 1GA, United Kingdom.
(2)Department of Biochemistry and Cambridge Systems Biology Centre, University of
Cambridge, Cambridge CB2 1GA, United Kingdom; The Francis Crick Institute, Mill
Hill Laboratory, The Ridgeway, Mill Hill, London NW7 1AA, United Kingdom.
Electronic address: [email protected].

The polyamines (PAs) spermidine, spermine, putrescine and cadaverine are an
essential class of metabolites found throughout all kingdoms of life. In this
comprehensive review, we discuss their metabolism, their various intracellular
functions and their unusual and conserved regulatory features. These include the
regulation of translation via upstream open reading frames, the over-reading of
stop codons via ribosomal frameshifting, the existence of an antizyme and an
antizyme inhibitor, ubiquitin-independent proteasomal degradation, a complex
bi-directional membrane transport system and a unique posttranslational
modification-hypusination-that is believed to occur on a single protein only
(eIF-5A). Many of these features are broadly conserved indicating that PA
metabolism is both concentration critical and evolutionary ancient. When PA
metabolism is disrupted, a plethora of cellular processes are affected, including
transcription, translation, gene expression regulation, autophagy and stress
resistance. As a result, the role of PAs has been associated with cell growth,
aging, memory performance, neurodegenerative diseases, metabolic disorders and
cancer. Despite comprehensive studies addressing PAs, a unifying concept to
interpret their molecular role is missing. The precise biochemical function of
polyamines is thus one of the remaining mysteries of molecular cell biology.



18. Am J Clin Nutr. 2015 Aug;102(2):411-9. doi: 10.3945/ajcn.114.103895. Epub 2015
Jul 1.

Dietary polyamine intake and colorectal cancer risk in postmenopausal women.

Vargas AJ(1), Ashbeck EL(2), Wertheim BC(2), Wallace RB(3), Neuhouser ML(4),
Thomson CA(5), Thompson PA(6).

Author information:
(1)Departments of Nutritional Sciences, [email protected].
(2)The University of Arizona Cancer Center, Tucson, AZ;
(3)University of Iowa College of Public Health, University of Iowa, Iowa City,
IA; and.
(4)Cancer Prevention Program, Fred Hutchinson Cancer Research Center, Seattle,
WA.
(5)Departments of Nutritional Sciences, College of Public Health, The University
of Arizona, Tucson, AZ; The University of Arizona Cancer Center, Tucson, AZ;
(6)Departments of Nutritional Sciences, Molecular and Cellular Biology, and The
University of Arizona Cancer Center, Tucson, AZ;

BACKGROUND: Putrescine, spermidine, and spermine (i.e., polyamines) are small
cationic amines synthesized by cells or acquired from the diet or gut bacteria.
Polyamines are required for both normal and colorectal cancer (CRC) cell growth.
OBJECTIVE: We investigated the association between dietary polyamines and risk of
CRC incidence and mortality.
DESIGN: The study was a prospective analysis in 87,602 postmenopausal women in
the Women's Health Initiative Observational Study. Multivariate Cox regression
was used to calculate HRs and 95% CIs.
RESULTS: Total dietary polyamine intake (mean ± SD: 289.2 ± 127.4 μmol/d) was not
positively associated with CRC in fully adjusted models. Instead, intake ≥179.67
μmol/d was associated with reduced risk of CRC [HR (95% CI): 0.82 (0.68, 1.00),
0.81 (0.66, 0.99), 0.91 (0.74, 1.12), and 0.80 (0.62, 1.02) for quintiles 2-5,
respectively, compared with quintile 1]. Reduced risk was not significant across
all quintiles. Polyamines were not significantly associated with CRC-specific
mortality in fully adjusted models. When stratified by risk factors for CRC, only
body mass index (BMI) and fiber intake significantly modified the association
between polyamine intake and CRC. In women with BMI (in kg/m²) ≤25 or fiber
consumption above the median, polyamine intake was associated with significantly
lower risk of CRC.
CONCLUSIONS: No positive association between dietary polyamines and CRC or
CRC-specific mortality risk in women was observed. Instead, a protective effect
of dietary polyamines was suggested in women with some CRC risk-lowering
behaviors in particular. These results are consistent with emerging evidence that
exogenous polyamines may be beneficial in colon health and warrant additional
study.



19. Cell Metab. 2015 Jun 2;21(6):891-7. doi: 10.1016/j.cmet.2015.04.011. Epub 2015
May 7.

Metabolism links bacterial biofilms and colon carcinogenesis.

Johnson CH(1), Dejea CM(2), Edler D(3), Hoang LT(1), Santidrian AF(4), Felding
BH(4), Ivanisevic J(1), Cho K(5), Wick EC(6), Hechenbleikner EM(6), Uritboonthai
W(1), Goetz L(7), Casero RA Jr(8), Pardoll DM(8), White JR(9), Patti GJ(5), Sears
CL(10), Siuzdak G(11).

Author information:
(1)Scripps Center for Metabolomics and Mass Spectrometry, The Scripps Research
Institute, La Jolla, CA 92037, USA.
(2)Department of Molecular Microbiology and Immunology, Bloomberg School of
Public Health, Johns Hopkins Medical Institutions, Baltimore, MD 21218, USA.
(3)Center of Surgical Gastroenterology, Karolinska University Hospital, 171 77
Stockholm, Sweden.
(4)Departments of Chemical Physiology and Molecular and Experimental Medicine,
The Scripps Research Institute, La Jolla, CA 92037, USA.
(5)Departments of Chemistry, Genetics, and Medicine, Washington University School
of Medicine, St. Louis, MO 63110, USA.
(6)Department of Surgery, Johns Hopkins University School of Medicine, Johns
Hopkins Medical Institutions, Baltimore, MD 21218, USA.
(7)Department of Surgery, Scripps Clinic Medical Group, La Jolla, CA 92037, USA.
(8)Departments of Oncology and Medicine and the Sidney Kimmel Comprehensive
Cancer Center, Johns Hopkins School of Medicine, Johns Hopkins Medical
Institutions, Baltimore, MD 21218, USA.
(9)Resphera Biosciences, Baltimore, MD 21231, USA.
(10)Department of Molecular Microbiology and Immunology, Bloomberg School of
Public Health, Johns Hopkins Medical Institutions, Baltimore, MD 21218, USA;
Departments of Oncology and Medicine and the Sidney Kimmel Comprehensive Cancer
Center, Johns Hopkins School of Medicine, Johns Hopkins Medical Institutions,
Baltimore, MD 21218, USA. Electronic address: [email protected].
(11)Scripps Center for Metabolomics and Mass Spectrometry, The Scripps Research
Institute, La Jolla, CA 92037, USA. Electronic address: [email protected].

Bacterial biofilms in the colon alter the host tissue microenvironment. A role
for biofilms in colon cancer metabolism has been suggested but to date has not
been evaluated. Using metabolomics, we investigated the metabolic influence that
microbial biofilms have on colon tissues and the related occurrence of cancer.
Patient-matched colon cancers and histologically normal tissues, with or without
biofilms, were examined. We show the upregulation of polyamine metabolites in
tissues from cancer hosts with significant enhancement of N(1),
N(12)-diacetylspermine in both biofilm-positive cancer and normal tissues.
Antibiotic treatment, which cleared biofilms, decreased N(1),
N(12)-diacetylspermine levels to those seen in biofilm-negative tissues,
indicating that host cancer and bacterial biofilm structures contribute to the
polyamine metabolite pool. These results show that colonic mucosal biofilms alter
the cancer metabolome to produce a regulator of cellular proliferation and colon
cancer growth potentially affecting cancer development and progression.



20. Prostate. 2015 Aug 1;75(11):1150-9. doi: 10.1002/pros.22996. Epub 2015 Apr 20.

Expression of spermidine/spermine N(1) -acetyl transferase (SSAT) in human
prostate tissues is related to prostate cancer progression and metastasis.

Huang W(1), Eickhoff JC(2), Mehraein-Ghomi F(3), Church DR(3), Wilding G(3), Basu
HS(3).

Author information:
(1)Department of Pathology and Laboratory Medicine, University of Wisconsin
Carbone Cancer Center, University of Wisconsin-Madison, Madison, Wisconsin.
(2)Department of Biostatistics, University of Wisconsin Carbone Cancer Center,
University of Wisconsin-Madison, Madison, Wisconsin.
(3)Department of Medicine, University of Wisconsin Carbone Cancer Center,
University of Wisconsin-Madison, Madison, Wisconsin.

INTRODUCTION: Prostate cancer (PCa) in many patients remains indolent for the
rest of their lives, but in some patients, it progresses to lethal metastatic
disease. Gleason score is the current clinical method for PCa prognosis. It
cannot reliably identify aggressive PCa, when GS is ≤ 7. It is shown that
oxidative stress plays a key role in PCa progression. We have shown that in
cultured human PCa cells, an activation of spermidine/spermine N(1) -acetyl
transferase (SSAT; EC 2.3.1.57) enzyme initiates a polyamine oxidation pathway
and generates copious amounts of reactive oxygen species in polyamine-rich PCa
cells.
METHOD: We used RNA in situ hybridization and immunohistochemistry methods to
detect SSAT mRNA and protein expression in two tissue microarrays (TMA) created
from patient's prostate tissues. We analyzed 423 patient's prostate tissues in
the two TMAs.
RESULTS: Our data show that there is a significant increase in both SSAT mRNA and
the enzyme protein in the PCa cells as compared to their benign counterpart. This
increase is even more pronounced in metastatic PCa tissues as compared to the PCa
localized in the prostate. In the prostatectomy tissues from early-stage
patients, the SSAT protein level is also high in the tissues obtained from the
patients who ultimately progress to advanced metastatic disease.
DISCUSSION: Based on these results combined with published data from our and
other laboratories, we propose an activation of an autocrine feed-forward loop of
PCa cell proliferation in the absence of androgen as a possible mechanism of
castrate-resistant prostate cancer growth.

 

[Wagner83]

me:

Hi Ray,
I recently discovered thymoquinone, and heard you talked about it some time ago. It has many interesting properties including crossing the BBB with ease. Do you have any idea what a safe dose would be?

Spoiler: References

Thymoquinone: an emerging natural drug with a wide range of medical applications

In this review we aimed at summarizing the most recent investigations related to a few and most important effects of thymoquinone. It is concluded that thymoquinone has evidently proved its activity as hepatoprotective, anti-inflammatory, antioxidant, cytotoxic and anti-cancer chemical, with specific mechanisms of action, which provide support to consider this compound as an emerging drug.

n traditional medicine, N. sativa has been used in different forms to treat many diseases including asthma, hypertension, diabetes, inflammation, cough, bronchitis, headache, eczema, fever, dizziness and influenza (1, 2).

Thymoquinone: An edible redox-active quinone for the pharmacotherapy of neurodegenerative conditions and glial brain tumors. A short review - ScienceDirect

Recently emerging studies conducted with animal models suggest that thymoquinone – bearing a very simple molecular structure – significantly crosses the blood brain barrier and exerts neuromodulatory activities. Indeed, in animal studies, the following actions of thymoquinone were demonstrated: 1—Protection against ischemic brain damage. 2—Reduction of epileptic seizures and associated cerebral oxidative injury. 3—Reduction of morphine tolerance and associated oxidative brain damage. 4—Anxiolytic effects and reduction of immobility stress-associated cerebral oxidative injury. 5—Reduction of diabetes-induced cerebral oxidative stress, 6—Reduction of cerebral oxidative injuries induced by noxious exposures including toluene, lead and ionizing radiation. Substantial in vitro data suggest that thymoquinone may be beneficial in treatment of glial tumors. However, there is no clinical study investigating its antitumor effects. In fact, thymoquinone suppresses growth and invasion, and induces apoptosis of glial tumor cells via degrading tubulins and inhibiting 20S proteasome, telomerase, autophagy, FAK and metalloproteinases.

Ray:

Quinones in the presence of reductants and catalysts can become semiquinones in a cycle that produces reactive oxygen species, so I think it’s risky except for treating an existing cancer.

Food Chem Toxicol. 2009 Jan;47(1):129-33.
In vitro toxicological properties of thymoquinone.
Khader M, Bresgen N, Eckl PM.
Nigella sativa has been traditionally used for the treatment of inflammations, liver disorders, and arthritis. Experimentally, it has been demonstrated that N. sativa extracts and the main constituent of their volatile oil, thymoquinone, possess antioxidant, anti-inflammatory and hepato-protective properties. To further evaluate the toxicological properties in a metabolically competent cellular system, thymoquinone was applied to primary rat hepatocyte cultures, and both cyto- and genotoxic effects were tested. Mitotic indices and the rates of apoptoses and necroses were determined as endpoints of cytotoxicity, while chromosomal aberrations and micronucleated cells served as endpoints of genotoxicity. In this approach thymoquinone demonstrated cyto- and genotoxic effects in a concentration dependent manner: it induced significant anti-proliferative effects at 20 microM and acute cytotoxicity at higher concentrations. Thymoquinone significantly increased the rates of necrotic cells at concentrations between 2.5 and 20 microM. Furthermore, it induced significant genotoxicity at concentrations > or =1.25 microM. These observations support the previous finding that thymoquinone causes glutathione depletion and liver damage, but contradict the reports indicating antioxidant and anti-clastogenic effects. Thymoquinone might be metabolised to reactive species and increase oxidative stress, which contributes to the depletion of antioxidant enzymes and damage to DNA in hepatocytes treated with high thymoquinone concentrations.

 

[DaveFoster]

I'm taking lamotrigine for bipolar symptoms, and I'm just curious as to your thoughts regarding the potential for things that lower anxiety (such as anticonvulsants) in the context of a high adrenaline response to thyroid. I don't think they improve glucose metabolism, but do you think the anxiolysis can be helpful in this context?

Dr. Peat: "There are safe, physiological methods that work better. I think chlorinated hydrocarbons should be avoided as far as possible.

Spoiler: Dangers of Lamotrigine

Wikipedia: Side effects
Lamotrigine prescribing information has a black box warning about life-threatening skin reactions, including Stevens–Johnson syndrome (SJS), DRESS syndrome and toxic epidermal necrolysis (TEN).[39] The manufacturer states that nearly all cases appear in the first two to eight weeks of therapy,[39] or if the medication is suddenly stopped then resumed at the normal dosage.[citation needed] Patients should seek medical attention for any unexpected skin rash, as its presence is an indication of a possible serious or even deadly side-effect of the drug."

 

[Wagner83]

Follow up on the mail I posted 2 posts above:

me:
Thanks a lot!

Does that mean you are against supplementing vitamin K2, emodin or cascara (for digestion etc..) and drinking pau d'arco tea on a regular basis? Do you have any idea what a good dose would be for cancer if one were to use thymoquinone?

Ray:

Menadione, K3, more water soluble, is used with vitamin C to treat cancer, creating free radicals similar to radiation. The reductive cytoplasm of cancer cells maintains the toxic effect. The oil soluble quinones are less reactive in the body, especially when they are in an oxidizing intracellular environment, which they help to stabilize. I would be inclined to use quinones other than thymoquinone for cancer, along with activated carbonyls such as fisetin.

 

[raypeatclips]

Question about vitamin b supplements.

You should be able to tell quickly whether it’s benefitting you. I just heard from another person, 85, who has used supplements for many years, with hypertension, swollen legs, and typical degenerative diseases, who recently stopped all supplements, and his blood pressure returned to what it was 30 years ago, the other symptoms completely disappeared. It isn’t at all rare for people to have chronic disease cause by supplements.

 

[paymanz]

On the safety and usefulness of molecular hydrogen therapy:

It’s safe, and in some situations it seems to have good effects.

Asked him on the safety of denatonium benzoate containing ethanol,as a solvent:

That wouldn’t be harmful.

 

[DaveFoster]

With regards to a 1:1 T3 to T4 ratio you've mentioned for those with impaired livers, do you know of any lab values that would indicate too much T3 in their thyroid ratio and would suggest an increase in T4?

Dr. Peat: "No, that can happen when stress increases reverse T3."

Okay, so I suppose it's [the need for more T4 is] only reliant upon negative symptoms such as arrhythmias (such as when you took only T3).

Do you think the [tattoo] ink's safe or do you think it'll cause problems over the long-term?

Dr. Peat: "I don’t know of any safe ink, and people’s reactions vary; the body can begin to react long after it was done."

...if one's sensitized to adrenaline, supplemented thyroid should be increased slowly. Once the adrenaline comes down, can the thyroid dose be adjusted more drastically (say in the months between winter and summer) without any fear of the same adrenaline surges?

Dr. Peat: "Yes."

 

[goodandevil]

On nitrous oxide:

Q: "is n2o inhalation harmful? The most i've read about is b12 deficiency." NB: i think he mentioned in an article that anesthetic gases like n2o can order the cell.

Ray: "Some of it breaks down to form NO; it isn’t the most harmful anesthetic, but it can be slightly harmful."

 

[goodandevil]

On the anti-inflammatory mechanism of cortisol, and the similarity of cortisol to progesterone with regards to protecting vascular endothelium:

Q: "How does "cortisol protect against shock and stress partly by maintaining the resistance and integrity of the capillaries"?

R: "Progesterone and cortisol both prevent leakiness of capillaries and are antiinflammatory, though they are antagonists in other situations. They both stabilize mast cells, decreasing histamine and serotonin, and inhibit phospholipase and prostaglandin formation and release of various inflammatory cytokines, and protect the glycocalyx."

 

[pepsi]

I emailed Dr Ray Peat about best way to deal with or treat stage 4 incurable brain cancer - GBM - glioblastoma multiforme.

He included a long, long list of references. I included them since someone might find them interesting to read.

Dr. Ray Peats email reply:

Tumors have multiple causes, so I think it’s best to use things that are known to be protective against known causes. I think cancers are continually produced and maintained by general metabolic conditions, and if those are ignored while trying to kill all of the abnormal cells, those cells release signals to recruit replacement cells, which, encountering the same or worse metabolic conditions, renew the tumor. Known causes involve inflammatory, excitatory processes, and when those are eliminated, tumors tend to disintegrate, undergoing “apoptosis,” a form of cell death that doesn’t create new inflammation. Here are some articles describing the effects of some of these antiinflammatory, antiexcitatory, substances—aspirin, caffeine, tetracylines, antihistamine/antiserotonin/anti-nitric oxide agents, progesterone, pregnenolone, antagonists of excitotoxicity, estrogen antagonists, vitamins D, E, and K, etc. Vitamin E (500 mg mixed tocopherols) and aspirin are things that the medical authorities might not object to.


1. Biomed Rep. 2016 Apr;4(4):444-448.
Cyclooxygenase inhibitor induces the upregulation of connexin-43 expression in C6
glioma cells.
Qin LJ(1), Jia YS(2), Zhang YB(1), Wang YH(1).
(1)Department of Physiology, School of Basic Medical Sciences, North China
University of Science and Technology, Tangshan, Hebei 063000, P.R. China.
(2)College of Traditional Chinese Medicine, North China University of Science and
Technology, Tangshan, Hebei 063000, P.R. China.
The present study was performed to determine whether aspirin, a cyclooxygenase
(COX) inhibitor, has an effect on the expression of connexin 43 (Cx43) in C6
glioma cells. Using an in vitro glioma invasion model, the expression of Cx43
protein in C6 cells was significantly increased following aspirin treatment at a
dose of 8 mmol/l for 30, 60 and 120 min via western blot analysis. The peak value
of the Cx43 expression was observed in C6 cells after 120 min of aspirin
treatment, which was significantly reduced by prostaglandin E2 (PGE2). In
addition, aspirin also significantly increased the gap junction intercellular
communication (GJIC) activity and reduced glioma invasion, which was induced by
PGE2. This led to the conclusion that the aspirin-induced glioma invasion
decrease may be associated with the increased expression of Cx43 protein and
formation of GJIC.
2. Eur J Cancer Prev. 2013 Nov;22(6):585-95.
Overexpression of S100A9 in human glioma and in-vitro inhibition by aspirin.
Huang N(1), Chen S, Deng J, Huang Q, Liao P, Wang F, Cheng Y.
(1)aDepartment of Neurosurgery, The Second Affiliated Hospital of Chongqing
Medical University bDepartment of Neurosurgery, The First Affiliated Hospital of
Chongqing Medical University cDepartment of Biomedical Engineering dInstitute of
Life Sciences, Chongqing Medical University, Chongqing, China.
Our previous work has shown that S100A9 promotes the growth of glioma cells. The
aim of this study was to investigate S100A9 expression in glioma cells and to
explore the potential of NSAIDs in the inhibition of S100A9. The levels of S100A9
were analyzed in five normal human brain tissues and 109 astrocytomas by
immunohistochemical analysis. In addition, S100A9 levels were detected in normal
human astrocytes, glioma cell lines, and six pairs of matched astrocytoma tissues
by reverse transcription-PCR or western blotting analysis. After treatment with
4, 8, and 16 mmol/l aspirin, cell viability, early apoptosis rate, and S100A9
levels were quantified. Cell viability and the changes in S100A9 levels were also
examined in glioma cells exposed to a cyclooxygenase-2 inhibitor, NS-398, alone
and in combination with prostaglandin E2. We found that S100A9 was upregulated in
astrocytomas and was significantly (P<0.05) correlated with histologic grades.
S100A9 protein levels were also elevated in six astrocytomas compared with
matched adjacent noncancerous tissues. Both S100A9 mRNA and protein levels were
higher in glioma cell lines than in normal human astrocytes (P<0.05). Aspirin
treatment inhibited cell proliferation and caused early apoptosis in glioma,
coupled with reduced S100A9 levels. Treatment with NS-398 decreased cell growth
and expression of S100A9 in glioma cells; these effects were partially reversed
by exogenous prostaglandin E2. These results suggest overexpression of S100A9 in
glioma cells. Aspirin may be a novel candidate for targeted prevention of S100A9
overexpression in glioma cells.
3. CNS Neurosci Ther. 2013 Feb;19(2):98-108.
Aspirin-/TMZ-coloaded microspheres exert synergistic antiglioma efficacy via
inhibition of β-catenin transactivation.
Shi ZD(1), Qian XM, Liu CY, Han L, Zhang KL, Chen LY, Zhang JX, Pu PY, Yuan XB,
Kang CS; Chinese Glioma Cooperative Group (CGCG).
(1)Laboratory of Neuro-Oncology, Department of Neurosurgery, Tianjin Neurological
Institute, Tianjin Medical University General Hospital, China.
BACKGROUND AND AIMS: Currently temozolomide (TMZ) as a potent agent is widely
used to treat the glioblastoma multiforme (GBM), whereas recurrence due to
intrinsic or acquired therapeutic resistance often occurs. Combination
chemotherapy with TMZ may be a promising therapeutic strategy to improve
treatment efficacy.
METHODS: Aspirin, TMZ, and aspirin-/TMZ-coloaded poly (L-lactide-co-glycolide)
(PLGA) microspheres were prepared by spray drying, and cytotoxicities of
glioblastoma cells were measured.
RESULTS: Aspirin microsphere treatment induced slight apoptosis and modestly
inhibited proliferation of LN229 and U87 cells in vitro and in vivo through
inhibition of β-catenin transactivation. However, aspirin-/TMZ-coloaded
microspheres presented synergistic antitumor efficacy compared with single
TMZ-loaded microspheres. Aspirin/TMZ microspheres induced more apoptosis and
repressed proliferation of LN229 and U87 cells. Corresponding to inhibition of
β-catenin signaling, β-catenin/TCF4 transcriptional activity and STAT3 luciferase
activity were strongly suppressed, and downstream targets expression was
decreased. Furthermore, aspirin/TMZ microsphere intratumoral injection
downregulated the expression of β-catenin, TCF4, pAKT, pSTAT3, and PCNA and
delayed tumor growth in nude mice harboring subcutaneous LN229 xenografts.
CONCLUSIONS: Aspirin sensitized TMZ chemotherapy efficacy through inhibition of
β-catenin transactivation; furthermore, the coloaded microspheres achieved a
sustained release action to reduce the TMZ dosage, offering the potential for
improved treatment of glioblastomas.
4. J Neurosurg. 2011 Oct;115(4):780-8.
Antitumor effect of aspirin in glioblastoma cells by modulation of
β-catenin/T-cell factor-mediated transcriptional activity.
Lan F(1), Yue X, Han L, Yuan X, Shi Z, Huang K, Yang Y, Zou J, Zhang J, Jiang T,
Pu P, Kang C.
(1)Laboratory of Neuro-Oncology, Tianjin Neurological Institute, Tianjin Medical
University General Hospital, Taijin, People’s Republic of China.
OBJECT: The goal in this study was to investigate the antitumor effect of aspirin
in glioblastoma cells and the molecular mechanism involved in its antineoplastic
activities.
METHODS: The authors used the
3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide method, flow
cytometry, the annexin V method, and Transwell cell invasion test to detect the
proliferation and invasive activity of U87 and A172 glioma cells before and after
being treated with aspirin. To determine the effects of aspirin on
β-catenin/T-cell factor (TCF) transcription activity, reporter constructs
containing 3 repeats of the wild-type (TOPflash) or mutant (FOPflash) TCF-binding
sites were used. Reverse transcriptase polymerase chain reaction and Western blot
analyses were used to detect the expression of multiple β-catenin/TCF target
genes following aspirin treatment.
RESULTS: The transcriptional activity of the β-catenin/TCF complex was strongly
inhibited by aspirin. Increasing the concentration of aspirin resulted in
decreased expression of c-myc, cyclin D1, and fra-1 mRNA and protein in U87 and
A172 cells in a dose-dependent manner. Aspirin inhibited glioma cell
proliferation and invasive ability, and induced apoptotic cell death.
CONCLUSIONS: The results suggest that aspirin is a potent antitumor agent, and
that it exerts its antineoplastic action by inhibition of the β-catenin/TCF
signaling pathway in glioma cells.
5. Neurol Res. 2003 Jun;25(4):370-6.
Aspirin and indomethacin exhibit antiproliferative effects and induce apoptosis
in T98G human glioblastoma cells.
Amin R(1), Kamitani H, Sultana H, Taniura S, Islam A, Sho A, Ishibashi M, Eling
TE, Watanabe T.
(1)Department of Neurosurgery, Institute of Neurological Sciences, Faculty of
Medicine, Tottori University School of Medicine, 36-1 Nishi-cho, Yonago, Tottori
683-8504, Japan.
The in vitro antiproliferative and apoptosis inducing properties of the
nonsteroidal anti-inflammatory drugs (NSAIDs) like acetyl salicylic acid
(aspirin) and indomethacin were investigated in T98G human glioblastoma cells to
explore their potential role in the chemoprevention of human glioma. The
biological effects induced by aspirin and indomethacin on T98G cells, in which
the expression of cyclooxygenase-1 (COX-1) and cyclooxygenase-2 (COX-2) were
confirmed by RT-PCR and immunostaining, were investigated by studying cell
proliferation and apoptosis assays. The antiproliferative effects occurred in a
dose- and time-dependent manner on T98G cells by the treatment with 0.1 -2 mM
aspirin and 25-100 microM indomethacin. Moreover, aspirin displayed the greatest
growth inhibition within 24 h. Approximately 90% growth inhibition occurred
following treatment either with 2 mM aspirin or 100 microM indomethacin by 72 h
and induction of apoptosis was confirmed by DNA laddering and TUNEL assay. Our in
vitro findings indicate that aspirin and indomethacin have an antiproliferative
effect on T98G human glioblastoma cells at toxic concentrations.
Mol Cells. 2011 Mar;31(3):275-9.
Caffeine inhibits cell proliferation and regulates PKA/GSK3β pathways in U87MG
human glioma cells.
Ku BM(1), Lee YK, Jeong JY, Ryu J, Choi J, Kim JS, Cho YW, Roh GS, Kim HJ, Cho
GJ, Choi WS, Kang SS.
(1)Department of Anatomy and Neurobiology, Institute of Health Science, School of
Medicine, Gyeongsang National University, Jinju, 660-751, Korea.
Caffeine is the most commonly ingested methylxanthine and has anti-cancer effects
in several types of cancer. In this study, we examined the anti-cancer effects of
caffeine on gliomas, both in vitro and in vivo. In vitro, caffeine treatment
reduced glioma cell proliferation through G(0)/G(1)-phase cell cycle arrest by
suppressing Rb phosphorylation. In addition, caffeine induced apoptosis through
caspase-3 activation and poly(ADP-ribose) polymerase (PARP) cleavage. Caffeine
also phosphorylated serine 9 of glycogen synthase kinase 3 beta (GSK3β).
Pretreatment with H89, a pharmacological inhibitor of protein kinase A (PKA), was
able to antagonize caffeine-induced GSK3β(ser9) phosphorylation, suggesting that
the mechanism might involve a cAMP-dependent PKA-dependent pathway. In vivo,
caffeine-treated tumors exhibited reduced proliferation and increased apoptosis
compared with vehicle-treated tumors. These results suggest that caffeine induces
cell cycle arrest and caspase-dependent cell death in glioma cells, supporting
its potential use in chemotherapeutic options for malignant gliomas.
Nutr Cancer. 2014;66(3):419-23.
Theobromine, the primary methylxanthine found in Theobroma cacao, prevents
malignant glioblastoma proliferation by negatively regulating
phosphodiesterase-4, extracellular signal-regulated kinase, Akt/mammalian target
of rapamycin kinase, and nuclear factor-kappa B.
Sugimoto N(1), Miwa S, Hitomi Y, Nakamura H, Tsuchiya H, Yachie A.
(1)a Department of Physiology, Graduate School of Medical Science , Kanazawa
University , Kanazawa , Japan.
Theobromine, a caffeine derivative, is the primary methylxanthine produced by
Theobroma cacao. We previously showed that methylxanthines, including caffeine
and theophylline, have antitumor and antiinflammatory effects, which are in part
mediated by their inhibition of phosphodiesterase (PDE). A member of the PDE
family, PDE4, is widely expressed in and promotes the growth of glioblastoma, the
most common type of brain tumor. The purpose of this study was to determine
whether theobromine could exert growth inhibitory effects on U87-MG, a cell line
derived from human malignant glioma. We show that theobromine treatment elevates
intracellular cAMP levels and increases the activity of p38 mitogen-activated
protein kinase and c-Jun N-terminal kinase, whereas it attenuates p44/42
extracellular signal-regulated kinase activity and the Akt/mammalian target of
rapamycin kinase and nuclear factor-kappa B signal pathways. It also inhibits
cell proliferation. These results suggest that foods and beverages containing
cocoa bean extracts, including theobromine, might be extremely effective in
preventing human glioblastoma.
Cancer Res. 2010 Feb 1;70(3):1173-83.
Caffeine-mediated inhibition of calcium release channel inositol
1,4,5-trisphosphate receptor subtype 3 blocks glioblastoma invasion and extends
survival.
Kang SS(1), Han KS, Ku BM, Lee YK, Hong J, Shin HY, Almonte AG, Woo DH, Brat DJ,
Hwang EM, Yoo SH, Chung CK, Park SH, Paek SH, Roh EJ, Lee SJ, Park JY, Traynelis
SF, Lee CJ.
(1)Center for Neural Science, Future Fusion Technology Laboratory, Seoul National
University, Seoul, Republic of Korea.
Calcium signaling is important in many signaling processes in cancer cell
proliferation and motility including in deadly glioblastomas of the brain that
aggressively invade neighboring tissue. We hypothesized that disturbing Ca(2+)
signaling pathways might decrease the invasive behavior of giloblastoma,
extending survival. Evaluating a panel of small-molecule modulators of Ca(2+)
signaling, we identified caffeine as an inhibitor of glioblastoma cell motility.
Caffeine, which is known to activate ryanodine receptors, paradoxically inhibits
Ca(2+) increase by inositol 1,4,5-trisphospate receptor subtype 3 (IP(3)R3), the
expression of which is increased in glioblastoma cells. Consequently, by
inhibiting IP(3)R3-mediated Ca(2+) release, caffeine inhibited migration of
glioblastoma cells in various in vitro assays. Consistent with these effects,
caffeine greatly increased mean survival in a mouse xenograft model of
glioblastoma. These findings suggest IP(3)R3 as a novel therapeutic target and
identify caffeine as a possible adjunct therapy to slow invasive growth of
glioblastoma.
Mol Cancer Ther. 2017 Jan;16(1):217-227.
Trifluoperazine, a Well-Known Antipsychotic, Inhibits Glioblastoma Invasion by
Binding to Calmodulin and Disinhibiting Calcium Release Channel IP3R.
Kang S(1), Hong J(2), Lee JM(2)(3), Moon HE(4), Jeon B(5)(6), Choi J(7), Yoon
NA(1), Paek SH(4), Roh EJ(5)(6), Lee CJ(8)(3)(9), Kang SS(10).
(1)Department of Anatomy and Convergence Medical Science, Institute of Health
Sciences, School of Medicine, Gyeongsang National University, Jinju, Republic of
Korea.
(2)Center for Neuroscience and Functional Connectomics, Brain Science Institute,
Korea Institute of Science and Technology, Seoul, Republic of Korea.
(3)KU-KIST Graduate School of Converging Science and Technology, Korea
University, Seoul, Republic of Korea.
(4)Department of Neurosurgery, Seoul National University College of Medicine,
Seoul, Republic of Korea.
(5)Chemical Kinomics Research Center, Korea Institute of Science and Technology,
Seoul, Republic of Korea.
(6)Department of Biological Chemistry, Korea University of Science and
Technology, Daejeon, Republic of Korea.
(7)Gyeongnam Department of Environmental Toxicology and Chemistry, Korea
Institute of Toxicology, Jinju, Republic of Korea.
(8)Center for Neuroscience and Functional Connectomics, Brain Science Institute,
Korea Institute of Science and Technology, Seoul, Republic of Korea.
[email protected] [email protected].
(9)Neuroscience Program, University of Science and Technology, Daejeon, Republic
of Korea.
(10)Department of Anatomy and Convergence Medical Science, Institute of Health
Sciences, School of Medicine, Gyeongsang National University, Jinju, Republic of
Korea. [email protected] [email protected].
Calcium (Ca(2+)) signaling is an important signaling process, implicated in
cancer cell proliferation and motility of the deadly glioblastomas that
aggressively invade neighboring brain tissue. We have previously demonstrated
that caffeine blocks glioblastoma invasion and extends survival by inhibiting
Ca(2+) release channel inositol 1,4,5-trisphosphate receptor (IP3R) subtype 3.
Trifluoperazine (TFP) is an FDA-approved antipsychotic drug for schizophrenia.
Interestingly, TFP has been recently reported to show a strong anticancer effect
on lung cancer, hepatocellular carcinoma, and T-cell lymphoma. However, the
possible anticancer effect of TFP on glioblastoma has not been tested. Here, we
report that TFP potently suppresses proliferation, motility, and invasion of
glioblastoma cells in vitro, and tumor growth in in vivo xenograft mouse model.
Unlike caffeine, TFP triggers massive and irreversible release of Ca(2+) from
intracellular stores by IP3R subtype 1 and 2 by directly interacting at the
TFP-binding site of a Ca(2+)-binding protein, calmodulin subtype 2 (CaM2). TFP
binding to CaM2 causes a dissociation of CaM2 from IP3R and subsequent opening of
IP3R. Compared with the control neural stem cells, various glioblastoma cell
lines showed enhanced expression of CaM2 and thus enhanced sensitivity to TFP. On
the basis of these findings, we propose TFP as a potential therapeutic drug for
glioblastoma by aberrantly and irreversibly increasing Ca(2+) in glioblastoma
cells. Mol Cancer Ther; 16(1); 217-27. ©2016 AACR.
1. Med Hypotheses. 2009 Jun;72(6):629-30.
Evo-Devo and the evolution of cancer: a hypothesis for metamorphic therapies for
the cancers of prolactin-influenced tumourigenesis: with special reference to
glioblastoma multiforme (GBM).
Pearson RD(1).
(1)Gerstein Science Information Centre, University of Toronto, 7 King's College
Circle, Toronto, Ontario, Canada. [email protected]
Recalling the remarkable developmental similarities between cancer cells and
embryonic tissues, this paper argues that, by the process of retrodifferentiation
and heterochronization, stem cells that have become neoplastic could be said to
have undergone "cellular heterochrony." It theorizes, therefore, that hormones
are the major factor in the non-random regulation of cellular heterochrony in
tumourigenesis. Two recent articles confirm that there is low thyroxine and high
prolactin in glioblastomas. Thyroxine metamorphoses vertebrates' tissues so as to
mature the tissues, e.g., in amphibian metamorphosis. In 1896, thyroxine (horse
thyroid extract) was the first successful hormonal product to be used against a
fulminating breast cancer. Recent work confirms the important role of prolactin
in the induction and progression of mammary, prostate and colorectal tumours.
Although the pituitary is the main source of prolactin in vertebrates, there is
also placental production of prolactin, and paracrine production of prolactin by
tumours themselves. Since tumours produce their own prolactin, shutting down the
pituitary source has not proven wholly successful. Research to find prolactin
receptor antagonists is ongoing. Therefore, prolactin inhibitors (dopamine
agonists), prolactin receptor antagonists, plus thyroxine comprise a plausible
metamorphic therapy for shrinking solid tumour mass. By contrast with
"differentiation" therapies currently sought by stem cell oncologists, this paper
advocates "metamorphic" therapies, to introduce hormonal oncological knowledge of
how to modulate signalling pathways that are aberrant in the stem cells that give
rise to tumours. Despite subtle differences in these signalling translation
pathways and cascades, strategies exist that will allow these evolved
populations, going back to their stem precursors, to "metamorphose" or perhaps
apoptotically cease proliferation.
2. J Neurophysiol. 2004 Mar;91(3):1203-16.
Effects of prolactin on ionic membrane conductances in the human malignant
astrocytoma cell line U87-MG.
Ducret T(1), Vacher AM, Vacher P.
(1)INSERM EMI 0347 Signalisation et Mécanismes Moléculaires de l'Apoptose, and
Laboratoire de Physiologie et Physiopathologie de la Signalisation Cellulaire,
CNRS UMR 5543, Université de Bordeaux 2, 33076 Bordeaux Cedex, France.
Prolactin (PRL) is involved in numerous biological processes in peripheral
tissues and the brain. Although numerous studies have been conducted to elucidate
the signal transduction pathways associated with the PRL receptor, very few have
examined the role of ion conductances in PRL actions. We used the patch-clamp
technique in "whole cell" configuration and microspectrofluorimetry to
investigate the effects of PRL on membrane ion conductances in the U87-MG human
malignant astrocytoma cell line, which naturally expresses the PRL receptor. We
found that a physiological concentration (4 nM) of PRL exerted a biphasic action
on membrane conductances. First, PRL activated a Ca(2+)-dependent K(+) current
that was sensitive to CTX and TEA. This current depended on PRL-induced Ca(2+)
mobilization, through a JAK2-dependent pathway from a thapsigargin- and
2-APB-sensitive Ca(2+) pool. Second, PRL also activated an inwardly directed
current, mainly due to the stimulation of calcium influx via nickel- and
2-APB-sensitive calcium channels. Both phases resulted in membrane
hyperpolarizations, mainly through the activation of Ca(2+)-dependent K(+)
channels. As shown by combined experiments (electrophysiology and
microspectrofluorimetry), the PRL-induced Ca(2+) influx increased with cell
membrane hyperpolarization and conversely decreased with cell membrane
depolarization. Thus PRL-induced membrane hyperpolarizations facilitated Ca(2+)
influx through voltage-independent Ca(2+) channels. Finally, PRL also activated a
DIDS-sensitive Cl(-) current, which may participate in the PRL-induced
hyperpolarization. These PRL-induced conductance activations are probably related
to the PRL proliferative effect we have already described in U87-MG cells.
3. Glia. 2002 May;38(3):200-14.
Effects of prolactin on intracellular calcium concentration and cell
proliferation in human glioma cells.
Ducret T(1), Boudina S, Sorin B, Vacher AM, Gourdou I, Liguoro D, Guerin J,
Bresson-Bepoldin L, Vacher P.
(1)Laboratoire de Neurophysiologie, Centre National de la Recherche Scientifique
UMR 5543, Université de Bordeaux 2, France.
Prolactin (PRL) has several physiological effects on peripheral tissues and the
brain. This hormone acts via its membrane receptor (PRL-R) to induce cell
differentiation or proliferation. Using reverse transcription-polymerase chain
reaction (RT-PCR) combined with Southern blot analysis, we detected PRL-R
transcripts in a human glioma cell line (U87-MG) and in primary cultured human
glioblastoma cells. These transcripts were deleted or not in their extracellular
domains. We examined the effects of PRL on intracellular free Ca2+ concentration
([Ca2+](i)) in these cells in order to improve our understanding of the PRL
transduction mechanism, which is still poorly documented. [Ca2+](i) was measured
by microspectrofluorimetry using indo-1 as the Ca2+ fluorescent probe.
Spatiotemporal aspects of PRL-induced Ca2+ signals were investigated using
high-speed fluo-3 confocal imaging. We found that physiological concentrations
(0.4-4 nM) of PRL-stimulated Ca2+ entry and intracellular Ca2+ mobilization via a
tyrosine kinase-dependent mechanism. The two types of Ca2+ responses observed
were distinguishable by their kinetics: one showing a slow (type I) and the other
a fast (type II) increase in [Ca2+](i). The amplitude of PRL-induced Ca2+
increases may be sufficient to provoke several physiological responses, such as
stimulating proliferation. Furthermore, PRL induced a dose-dependent increase in
[3H]thymidine incorporation levels and in cellular growth and survival, detected
by the MTT method. These data indicate that PRL induced mitogenesis of human
glioma cells.

5. J Neurochem. 1999 Dec;73(6):2272-7.
Prolactin induction of nitric oxide synthase in rat C6 glioma cells.
Raso GM(1), Meli R, Gualillo O, Pacilio M, Di Carlo R.
(1)Department of Experimental Pharmacology, University of Naples Frederico II,
Italy.
We have examined the neuroimmunoregulatory function of prolactin (PRL) on
astrocytic inducible nitric oxide synthase (iNOS) expression in the C6 glioma
cell line. After 24 h of PRL (5-100 nM) stimulation, a concentration-dependent
increase of NO release, evaluated as nitrite, was observed in C6 culture medium.
Moreover, both NO release and iNOS expression induced by interferon-gamma (250
U/ml) were enhanced by PRL (18-100 nM). PRL-induced NO release was inhibited by
dexamethasone, an inhibitor of de novo iNOS synthesis. We used erbstatin (5
microg/ml), a potent inhibitor of protein tyrosine kinases, to test whether these
proteins were required for signaling events evoked by PRL in these cells. This
inhibitor was able to inhibit completely the PRL-induced NO production and iNOS
expression. In conclusion, we provide evidence that NO production in glial cells
can be regulated not only by cytokines but also by neuroimmunoregulatory hormones
such as PRL, which is present in normal brain but may be elevated in several
pathological states.
4. J Neurosurg Sci. 2001 Jun;45(2):70-4.
Hyperprolactinaemia and prolactin binding in benign intracranial tumours.
Ciccarelli E(1), Razzore P, Gaia D, Todaro C, Longo A, Forni M, Ghè C, Camanni F,
Muccioli G, Faccani G, Lanotte MM.
(1)Department of Internal Medicine, Division of Endocrinology, University of
Turin, Turin, Italy.
BACKGROUND: Meningiomas have been found to have receptors for several hormones,
such as oestrogen, progesterone, somatostatin, dopamine and recently also for
prolactin.
METHODS: To investigate any possible role of prolactin in the growth of those
tumours we detected the presence of prolactin-receptors (PRL-R) in 22 meningiomas
and we correlated these data with PRL serum levels in patients before surgery. We
also studied 13 patients with schwannomas and 7 with other cerebral tumours (4
glioblastomas, 2 ependymomas and 1 astrocytoma).
RESULTS: Increased prolactin binding was present in 10 (45.4 percent
meningiomas, 9 (69.2 percent schwannomas and in the patient with astrocytoma.
The presence of high PRL levels was present in 6 (27.2 percent patients with
meningiomas, 8 (61.5 percent with schwannomas and in 3 (42.8 percent with
other tumours. No direct correlation was present between serum PRL levels and PRL
binding in all groups.
CONCLUSIONS: In conclusion we confirmed the presence of PRL receptors in patients
with meningiomas and we have also shown the presence of PRL receptors also in
schwannomas. Moreover increased serum PRL were shown in some patients with
different tumours of nervous tissue before surgery. Our data could suggest that
PRL might have a role in the growth of meningiomas and schwannomas.
1. Int J Physiol Pathophysiol Pharmacol. 2017 Apr 15;9(2):8-15. eCollection 2017.
Lidocaine suppresses glioma cell proliferation by inhibiting TRPM7 channels.
Leng T(1), Lin S(2), Xiong Z(1), Lin J(3).
(1)Neurobiology, Neuroscience Institute, Morehouse School of MedicineAtlanta GA
30329, USA.
(2)Guangzhou Cellproteck Pharmaceutical CO., Ltd3 Lanyue Road, Science City,
Guangzhou 510663, PR China.
(3)Department of Anesthesiology, Stony Brook University Health Sciences Center,
Stony BrookNY 11794-8480, USA.
BACKGROUND: Malignant glioma is the most common brain cancer with devastating
prognosis. Recurrence of malignant glioma following surgery is very common with
few preventive and therapeutic options. Novel targets and therapeutic agents are
constantly sought for better outcome. Our previous study established that
inhibition of transient receptor potential melastatin 7 (TRPM7) channels resulted
in significant decrease of human glioma cell growth and proliferation. As local
anesthetic lidocaine has been shown to inhibit TRPM7 currents, we hypothesize
that lidocaine may suppress glioma cell proliferation through TRPM7 channel
inhibition.
METHODS: TRPM7 currents were recorded in rat C6 glioma cells using the whole cell
patch clamp technique. Cell growth and proliferation were assessed under
microscopic examination and biochemical assays.
RESULTS: Lidocaine inhibits TRPM7-like currents in a dose-dependent and
reversible manner. At 1 and 3 mM, it inhibits ~30% and ~50% of TRPM7 currents. At
these concentrations, it is effective in inhibiting the proliferation of C6
cells. As expected, the TRPM7 inhibitors gadolinium and 2-Aminoethoxydiphenyl
borate have similar effects on TRPM7 currents and proliferation of C6 cells.
Similar to its effect on C6 cells, lidocaine inhibits the proliferation of A172
cells, a human glioblastoma cell line.
CONCLUSIONS: Lidocaine significantly inhibits the proliferation of glioma cells.
The effect of lidocaine is mediated, at least in part, by inhibiting TRPM7
channels.
Anesth Analg. 2016 Apr;122(4):1038-47.
Clinical Concentrations of Local Anesthetics Bupivacaine and Lidocaine Differentially Inhibit Human Kir2.x Inward Rectifier K+ Channels.
Nakahira K, Oshita K, Itoh M, Takano M, Sakaguchi Y, Ishihara K.
BACKGROUND:
Inward rectifier K channels of the Kir2.x subfamily are widely expressed in neuronal tissues, controlling neuronal excitability. Previous studies reported that local anesthetics (LAs) do not affect Kir2 channels. However, the effects have not been studied at large concentrations used in regional anesthesia.
METHODS:
This study used the patch-clamp technique to examine the effects of bupivacaine and lidocaine on Kir2.1, Kir2.2, and Kir2.3 channels expressed in human embryonic kidney 293 cells.
RESULTS:
When applied extracellularly in whole-cell recordings, both LAs inhibited Kir2.x currents in a voltage-independent manner. Inhibition with bupivacaine was slow and irreversible, whereas that with lidocaine was fast and reversible. Kir2.3 displayed a greater sensitivity to bupivacaine than Kir2.1 and Kir2.2 (50% inhibitory concentrations at approximately 5 minutes, 0.6 vs 8-10 mM), whereas their sensitivities to lidocaine were similar (50% inhibitory concentrations, 1.5-2.7 mM). Increases in the charged/neutral ratio of the LAs at an acidic extracellular pH attenuated their inhibitory effects, and a permanently charged lidocaine derivative QX-314 exhibited no effects when applied extracellularly. Inside-out experiments demonstrated that inhibition of Kir2.1 with cytoplasmic lidocaine and QX-314 was rapid and reversible, whereas that induced by bupivacaine was slow and irreversible. Furthermore, dose-inhibition relations for the charged form of bupivacaine and lidocaine obtained at different cytoplasmic pHs could be approximated by a single relation for each LA.
CONCLUSIONS:
The results indicate that both LAs at clinical concentrations equilibrated rapidly with the intracellular milieu, differentially inhibiting Kir2.x channel function from the cytoplasmic side.
Anesthesiology. 2001 Jun;94(6):1089-95.
Local anesthetic inhibition of voltage-activated potassium currents in rat dorsal root ganglion neurons.
Komai H, McDowell TS.
BACKGROUND:
Local anesthetic actions on the K+ channels of dorsal root ganglion (DRG) and dorsal horn neurons may modulate sensory blockade during neuraxial anesthesia. In dorsal horn neurons, local anesthetics are known to inhibit transient but not sustained K+ currents. The authors characterized the effects of local anesthetics on K+ currents of isolated DRG neurons.
METHODS:
The effects of lidocaine, bupivacaine, and tetracaine on K+ currents in isolated rat DRG neurons were measured with use of a whole cell patch clamp method. The currents measured were fast-inactivating transient current (I(Af)), slow-inactivating transient current (I(As)), and noninactivating sustained current (I(Kn)).
RESULTS:
One group of cells (type 1) expressed I(Af) and I(Kn). The other group (type 2) expressed I(As) and I(Kn). The diameter of type 2 cells was smaller than that of type 1 cells. Lidocaine and bupivacaine inhibited all three K+ currents. Tetracaine inhibited I(As) and I(Kn) but not I(Af) For bupivacaine, the concentration for half-maximal inhibition (IC50) of I(Kn) in type 2 cells was lower than that for I(Kn) in type 1 cells (57 vs. 121 microM). Similar results were obtained for tetracaine (0.6 vs. 1.9 mM) and for lidocaine (2.2 vs. 5.1 mM).
CONCLUSIONS:
Local anesthetics inhibited both transient and sustained K+ currents in DRG neurons. Because K+ current inhibition is known to potentiate local anesthetic-induced impulse inhibition, the lower IC50 for I(Kn) of small type 2 cells may reflect preferential inhibition of impulses in nociceptive neurons. The overall modulatory actions of local anesthetics probably are determined by their differential effects on presynaptic (DRG) and postsynaptic (dorsal horn neurons) K+ currents.

Arch Biochem Biophys. 1985 Mar;237(2):415-22.
Induction of intracellular acyl-CoA:cholesterol acyltransferase activity in
glioblastoma cells by lidocaine.
Jeng I, Klemm N, Bressie S, Samson L.
The perturbation of cellular cholesteryl ester biosynthesis in glioblastoma C-6
cells by lidocaine was investigated. Lidocaine specifically inhibited the
incorporation of radioactive oleic acid into cellular cholesteryl ester but had
no significant effect on the incorporation of oleic acid into
phosphatidylcholine. Oxygenated cholesterol-enhanced cholesteryl ester formation
was less sensitive to lidocaine inhibition. Several other local anesthetics were
compared. Lidocaine altered cholesteryl ester formation in time- and
dose-dependent manners. Lidocaine was a powerful inhibitor initially and its
potency declined with time. Lidocaine was capable of directly inhibiting
acyl-CoA:cholesterol acyltransferase (ACAT) activity in broken cell homogenates.
The lidocaine-mediated inhibition of cellular cholesteryl ester formation
triggered an enhanced intracellular ACAT activity that was not fully expressed in
the presence of lidocaine. The activation of ACAT activity by lidocaine might
represent a compensatory mechanism by which the inhibitory effect of lidocaine
was partially overcome with time.
Int J Neurosci. 2007 Oct;117(10):1465-80.
Medroxyprogesterone acetate induces c6 glioma chemosensitization via
antidepressant-like lysosomal phospholipidosis/myelinosis in vitro.
Altinoz MA(1), Gedikoglu G, Sav A, Ozcan E, Ozdilli K, Bilir A, Del Maestro RF.
(1)Institute of Health, Halic University, Istanbul, Turkey. [email protected]
The authors have previously shown that medroxyprogesterone acetate (MPA) inhibits
growth and increases drug sensitivity in C6 glioma with myeloid bodies. Myeloid
bodies can occur in cells either due to robust toxicity with mitochondrial
membrane disruption or due to milder events such as seen in
lysosomal-phospholipidosis. Exact patterns of myelinosis accompanying to MPA
chemo-sensitization is important, because uncoupling of nuclear versus
mitochondrial toxicity of anti-neoplastics by MPA would lead to safer employment
of glioma chemotherapy with reduced neurotoxicity. By monitoring and comparing
cell kinetics with fine structural features of cell death, the authors estimated
subcellular effects accompanying growth-inhibitory drug actions in C6 glioma. The
analysis revealed that MPA induced mainly lysosomal phospholipidosis, while
inhibiting clonogenicity alone and augmenting procarbazine efficacy. It induced
apoptosis in combination with cisplatin. It reduced mitochondrial-damage-based
early cytotoxicity of methotrexate, yet it did not hinder its anti-clonogenic
efficacy. Progesterone analogues - similar to antidepressants - inhibit
cholesterol esterification, and this efficacy relates with their P-glycoprotein
inhibition. Reducing esterification and plasma-membrane localization of
cholesterol may lead MPA induction of lysosomal phospholipidosis, growth
indolency, and drug sensitization in glioma.
Int J Dev Neurosci. 2001 Oct;19(6):541-7.
Medroxyprogesterone acetate alone or synergistic with chemotherapy suppresses
colony formation and DNA synthesis in C6 glioma in vitro.
Altinoz MA(1), Bilir A, Ozar E, Onar FD, Sav A.
(1)Histology and Embryology Department, Istanbul Medical School, 34390, Istanbul,
Turkey. [email protected]
We have studied the effects of medroxyprogesterone acetate (MPA) on C6 glioma
growth in vitro in order to prove the hypothesis that it could arrest growth and
induce drug sensitisation in a glial tumour as it does in breast cancer cells.
Plating, thymidine-labelling index, ultra-structure, and soft agar colony growth
were determined after incubation with MPA, and/or cisplatin, procarbazine and
methotrexate (MTX). MPA (microg/ml) reduced the thymidine-labelling index by 41
and 73% at 48 and 96 h, respectively, and decreased colony growth by 61%. Soft
agar colony inhibition by MPA was almost as potent as MTX (0.3 microg/ml), but
the latter drug showed very high cytotoxicity. Electron microscopy revealed that
in medroxyprogesterone treated cells myeloid bodies developed, but MTX treatment
caused mainly necrosis. Medroxyprogesterone increased procarbazine and
cisplatin-induced colony growth and S-phase inhibition, but reduced MTX-induced
thymidine-labelling inhibition. In conclusion, progesterone may inhibit growth
and sensitize to drugs.
Brain Res. 1988 Mar 8;443(1-2):338-44.
Memantine (1-amino-3,5-dimethyladamantane) blocks the serotonin-induced
depolarization response in a neuronal cell line.
Reiser G(1), Binmöller FJ, Koch R.
(1)Physiologisch-chemisches Institut der Universität Tübingen, F.R.G.
The influence of memantine on several properties of a neuronal cell line was
tested. The aim was to get some insight into possible mechanisms of action of
this drug which is therapeutically applicable in treatment of spasticity,
Parkinson's disease, and cerebral coma. In neuroblastoma X glioma hybrid cells,
memantine, at micromolar concentrations, blocked the depolarization induced by
iontophoretically applied serotonin (5-hydroxytryptamine, 5-HT). In the hybrid
cells, receptors of the 5-HT3 type mediated the depolarization, which was
frequently accompanied by a series of action potentials. The inhibition by
memantine of the serotonin response occurred fast and was completely reversible,
irrespective of whether the cell showed a stable membrane potential or
spontaneous action potentials. However, memantine did not alter spontaneous or
electrically evoked action potential activity in the hybrid cells, and apparently
did not block the underlying ionic conductances. Furthermore memantine did not
affect either the cation permeability activated by substance P in the hybrid
cells or the K+ channel triggered by bradykinin in a glioma cell line. Thus,
memantine appears specifically to suppress the ion channel opened by serotonin in
the hybrid cells. The interaction of memantine with serotonin receptors and the
associated ion channels reported here, might give an important clue, as to a site
of action of memantine in the nervous system.
J Neurosurg. 2014 Apr;120(4):854-63.
Efficacy of local polymer-based and systemic delivery of the anti-glutamatergic
agents riluzole and memantine in rat glioma models.
Yohay K(1), Tyler B, Weaver KD, Pardo AC, Gincel D, Blakeley J, Brem H, Rothstein
JD.
(1)Department of Pediatrics, Weill Cornell Medical College;
OBJECT: The poor outcome of malignant gliomas is largely due to local
invasiveness. Previous studies suggest that gliomas secrete excess glutamate and
destroy surrounding normal peritumoral brain by means of excitotoxic mechanisms.
In this study the authors assessed the effect on survival of 2 glutamate
modulators (riluzole and memantine) in rodent glioma models.
METHODS: In an in vitro growth inhibition assay, F98 and 9L cells were exposed to
riluzole and memantine. Mouse cerebellar organotypic cultures were implanted with
F98 glioma cells and treated with radiation, radiation + riluzole, or vehicle and
assessed for tumor growth. Safety and tolerability of intracranially implanted
riluzole and memantine CPP:SA polymers were tested in F344 rats. The efficacy of
these drugs was tested against the 9L model and riluzole was further tested with
and without radiation therapy (RT).
RESULTS: In vitro assays showed effective growth inhibition of both drugs on F98
and 9L cell lines. F98 organotypic cultures showed reduced growth of tumors
treated with radiation and riluzole in comparison with untreated cultures or
cultures treated with radiation or riluzole alone. Three separate efficacy
experiments all showed that localized delivery of riluzole or memantine is
efficacious against the 9L gliosarcoma tumor in vivo. Systemic riluzole
monotherapy was ineffective; however, riluzole given with RT resulted in improved
survival.
CONCLUSIONS: Riluzole and memantine can be safely and effectively delivered
intracranially via polymer in rat glioma models. Both drugs demonstrate efficacy
against the 9L gliosarcoma and F98 glioma in vitro and in vivo. Although systemic
riluzole proved ineffective in increasing survival, riluzole acted
synergistically with radiation and increased survival compared with RT or
riluzole alone.
Bull Cancer. 2013 Sep;100(9):829-35.
[Glutamate and malignant gliomas, from epilepsia to biological aggressiveness:
therapeutic implications].
[Article in French]
Blecic S(1), Rynkowski M, De Witte O, Lefranc F.
(1)Service de neurologie, Epicura, 7800 Ath, Belgique.
In this review article, we describe the unrecognized roles of glutamate and
glutamate receptors in malignant glioma biology. The neurotransmitter glutamate
released from malignant glioma cells in the extracellular matrix is responsible
for seizure induction and at higher concentration neuronal cell death. This
neuronal cell death will create vacated place for tumor growth. Glutamate also
stimulates the growth and the migration of glial tumor cells by means of the
activation of glutamate receptors on glioma cells in a paracrine and autocrine
manner. The multitude of effects of glutamate in glioma biology supports the
rationale for pharmacological targeting of glutamate receptors and transporters
in the adjuvant treatment of malignant gliomas in neurology and neuro-oncology.
Using the website www.clinicaltrials.gov/ as a reference - a service developed by
the National Library of Medicine for the National Health Institute in USA - we
have evoked the few clinical trials completed and currently ongoing with
therapies targeting the glutamate receptors.
Oncol Lett. 2014 Aug; 8(2): 645–650.
Pregnenolone, a cholesterol metabolite, induces glioma cell apoptosis via activating extrinsic and intrinsic apoptotic pathways
XIAO XIAO,1,* LIJUN CHEN,1,* YING OUYANG,2,* WENBO ZHU,1 PENGXIN QIU,1 XINWEN SU,1 YUNLING DOU,3 LIPENG TANG,1 MIN YAN,1 HAIPENG ZHANG,1 XIAOXIAO YANG,1 DONG XU,1 and GUANGMEI YAN1
Gliomas are one of the most common types of malignant tumors worldwide, however, an effective therapeutic strategy not yet been fully determined. The present study aimed to investigate the anti-glioma activity and underlying mechanisms of pregnenolone, which originates from cholesterol and is metabolized into important steroid hormones in the body. The results demonstrated that 100 μM pregnenolone induced a significant loss of cell viability in various malignant glioma cell lines. In the U-87 MG, LN-18 and C6 cell lines, the loss of cell viability resulted from cell apoptosis, which was evidenced by apoptotic nuclear morphology changes and caspase 3 activation. Moreover, the increased activities of caspase 8 and 9 strongly indicated that pregnenolone activated the extrinsic and intrinsic pathways of apoptosis. Additionally, glioma cell apoptosis was prevented by the general caspase inhibitor, Z-VAD-FMK. In the C6 cells, upregulation of Fas and Fas ligand triggered the activation of the extrinsic pathway, whereas knockdown of Fas significantly abrogated the cell apoptosis that was induced by pregnenolone. Furthermore, downregulation of the anti-apoptotic protein, B-cell lymphoma 2 and upregulation of pro-apoptotic proteins, such as Bax and Bak, activated the intrinsic pathway. In conclusion, pregnenolone induced glioma cell apoptosis in a caspase-dependent manner, which was mediated by activation of the extrinsic and intrinsic apoptotic pathways.
Introduction
Gliomas originate from glial cells and are the most common type of primary brain tumors accounting for 80% of all malignant primary brain tumors (1). According to the pathological and clinical criteria established by the World Health Organization, gliomas are classified as grades I–IV (2,3). Grade IV tumors, such as glioblastomas (GBMs), are the most devastating and aggressive form comprising >50% of gliomas, and have a poor prognosis (4). The current treatment standard of GBMs is surgical resection to a feasible extent, followed by radiotherapy and chemotherapy. Among the currently available chemotherapy agents, temozolomide is the most popular; doctors and patients favor it as it is administrated orally and efficiently crosses the blood-brain barrier (BBB) (5). However, 67.2–76% of patients are resistant to this agent and therefore do not benefit from it (6). Regardless of systemic therapeutic strategies, including surgery, temozolomide and radiotherapy, patient median survival is only 14.6 months and the five-year survival rate is ~9.8% (7,8). The poor prognosis of glioma fuels the requirement for identifying therapeutic agents with the merits of temozolomide, such as high lipophilicity and strong anti-glioma activity.
Steroid hormones are generally divided into the following five groups: Estrogens, androgens, progestogens, glucocorticoids and mineralocorticoids (9,10). The natural steroid hormones are predominantly synthesized from cholesterol in the gonads and adrenal glands, and readily diffuse through the cell membrane due to their lipophilic properties (11,12). Accumulating evidence has indicated that certain steroid hormones possess antitumor activities, such as the 17β-estradiol metabolite, 2-methoxyestradiol (2ME), which exerts the strongest activity. 2ME inhibits proliferation and induces apoptosis of various types of cancer, including gliomas, and breast and gastric cancer, independently of estrogen receptors α and β (13–15). However, findings from a clinical trial identified low oral bioavailability of 2ME, which prevents the transfer of this promising agent from bench to bed side (16). The example of 2ME implies the application potential of steroid hormones and provides the basis for the subsequent investigation of other endogenous steroid hormones, such as pregnenolone (Fig. 1).
The present study aimed to investigate the pharmacological effects of the endogenous steroid, pregnenolone, on GBM cells, and the mechanisms underlying its pro-apoptotic activity via the extrinsic and intrinsic apoptotic pathways. Pregnenolone may be a leading compound in the treatment of gliomas and may be modified and developed for clinical application.
J Neurosurg. 1995 Apr;82(4):635-40.
The role of minocycline in the treatment of intracranial 9L glioma.
Weingart JD(1), Sipos EP, Brem H.
(1)Department of Neurological Surgery, Johns Hopkins University School of
Medicine, Baltimore, Maryland.
This study was designed to explore the question of whether minocycline, a
semisynthetic tetracycline shown to inhibit tumor-induced angiogenesis, could
control the growth of the rat intracranial 9L gliosarcoma. Minocycline was tested
alone and in combination with 1,3-bis(2-chloroethyl)-1-nitrosourea (BCNU) in
vivo. Treatment was started at the time of intracranial implantation of 9L
gliosarcoma into male Fischer 344 rats, 5 days later, or after tumor resection.
Minocycline was delivered locally with a controlled-release polymer or
systemically by intraperitoneal injection. Systemic minocycline did not extend
survival time. Local treatment with minocycline by a controlled-release polymer
implanted at the time of tumor implantation extended median survival time by 530%
(p < 0.001) compared to treatment with empty polymer. When treatment was begun 5
days after tumor implantation, minocycline delivered locally or systemically had
no effect on survival. However, after tumor resection, treatment with locally
delivered minocycline resulted in a 43% increase in median survival time (p <
0.002) compared to treatment with empty polymer. Treatment with a combination of
minocycline delivered locally in a controlled-release polymer and systemic BCNU 5
days after tumor implantation resulted in a 93% extension of median survival time
compared to BCNU alone (p < 0.002). In contrast, treatment with a combination of
systemic minocycline and BCNU did not increase survival time compared to systemic
BCNU alone. These results demonstrate that minocycline affects tumor growth when
delivered locally and suggest that minocycline may be a clinically effective
modulator of intracranial tumor growth when used in combination with a
chemotherapeutic agent and surgical resection.
J Neurooncol. 2003 Sep;64(3):203-9.
Local delivery of minocycline and systemic BCNU have synergistic activity in the
treatment of intracranial glioma.
Frazier JL(1), Wang PP, Case D, Tyler BM, Pradilla G, Weingart JD, Brem H.
(1)Department of Neurological Surgery, Johns Hopkins University School of
Medicine, Baltimore, MD 21205, USA.
Minocycline, a tetracycline derivative, has been shown to inhibit tumor
angiogenesis through inhibitory effects on matrix metalloproteinases. Previous
studies have shown this agent to be effective against a rodent brain tumor model
when delivered intracranially and to potentiate the efficacy of standard
chemotherapeutic agents. In the present study, the in vivo efficacy of
intracranial minocycline delivered by a biodegradable controlled-release polymer
against rat intracranial 9L gliosarcoma was investigated to determine whether it
potentiates the effects of systemic 1,3-bis (2-chloroethyl)-1-nitrosourea (BCNU).
Minocycline was incorporated into the biodegradable polymer polyanhydride
poly[bis(p-carboxyphenoxy)propane-sebacic acid] (pCPP:SA) at a ratio of 50:50 by
weight. The release kinetics of minocycline from the polymer were assessed. For
the efficacy studies, female Fischer 344 rats were implanted with 9L glioma.
Treatment with minocycline delivered by the pCPP:SA polymer at the time of tumor
implantation resulted in 100% survival in contrast to untreated control animals
that died within 21 days. Treatment with the minocycline-polymer 5 days after
tumor implantation provided only modest increases in survival. The combination of
intracranial minocycline and systemic BCNU extended median survival by 82%
compared to BCNU alone (p < 0.0001) and 200% compared to no treatment (p <
0.004). We conclude that local intracranial delivery of minocycline from
biodegradable controlled-release polymers inhibits tumor growth and may have
clinical utility when combined with a chemotherapeutic agent.
Int J Cancer. 2016 Jun 1;138(11):2678-87.
Investigating the role of tumour cell derived iNOS on tumour growth and
vasculature in vivo using a tetracycline regulated expression system.
Papaevangelou E(1), Whitley GS(1), Johnstone AP(1), Robinson SP(2), Howe FA(1).
(1)Cardiovascular and Cell Sciences Research Institute, St. George's, University
of London, Cranmer Terrace, London, SW17 0RE, United Kingdom. (2)Division of
Radiotherapy and Imaging, The Institute of Cancer Research, Belmont, Sutton,
Surrey, SM2 5NG, United Kingdom.
Nitric oxide (NO) is a free radical signalling molecule involved in various
physiological and pathological processes, including cancer. Both tumouricidal and
tumour promoting effects have been attributed to NO, making its role in cancer
biology controversial and unclear. To investigate the specific role of
tumour-derived NO in vascular development, C6 glioma cells were genetically
modified to include a doxycycline regulated gene expression system that controls
the expression of an antisense RNA to inducible nitric oxide synthase (iNOS) to
manipulate endogenous iNOS expression. Xenografts of these cells were propagated
in the presence or absence of doxycycline. Susceptibility magnetic resonance
imaging (MRI), initially with a carbogen (95% O2 /5% CO2 ) breathing challenge
and subsequently an intravascular blood pool contrast agent, was used to assess
haemodynamic vasculature (ΔR2 *) and fractional blood volume (fBV), and
correlated with histopathological assessment of tumour vascular density,
maturation and function. Inhibition of NO production in C6 gliomas led to
significant growth delay and inhibition of vessel maturation. Parametric fBV maps
were used to identify vascularised regions from which the carbogen-induced ΔR2 *
was measured and found to be positively correlated with vessel maturation,
quantified ex vivo using fluorescence microscopy for endothelial and perivascular
cell staining. These data suggest that tumour-derived iNOS is an important
mediator of tumour growth and vessel maturation, hence a promising target for
anti-vascular cancer therapies. The combination of ΔR2 * response to carbogen and
fBV MRI can provide a marker of tumour vessel maturation that could be applied to
non-invasively monitor treatment response to iNOS inhibitors.
Cancer Lett. 1995 Jan 27;88(2):141-5.
Methylene blue induces cytotoxicity in human brain tumor cells.
Lee YS(1), Wurster RD.
(1)Department of Neurological Surgery, Loyola University Medical Center, Maywood,
IL 60153.
Methylene blue (MB), a known inhibitor of guanylyl cyclase, induced cytotoxicity
in SK-N-MC human neuroblastoma and U-373 MG human astrocytoma cells in a
dose-dependent manner. MB did not significantly alter cellular levels of cGMP in
both cells. 8-Br cGMP, a membrane-permeable analogue of cGMP, did not decrease
MB-induced cytotoxicity, indicating that cGMP may not be a major target of the
cytotoxic action of MB. However, hydroxyl radical scavengers or intracellular
Ca2+ modulators effectively blocked the MB-induced cytotoxicity. These results
suggest that hydroxyl radical and intracellular Ca2+ may have an important
involvement in the cytotoxic action of MB. These results further suggest that the
treatment with MB may be useful for the therapeutic applications of human brain
tumors.
Prog Neurobiol. 2015 Nov 18. pii: S0301-0082(15)30060-5.
Alternative mitochondrial electron transfer for the treatment of
neurodegenerative diseases and cancers: Methylene blue connects the dots.
Yang SH(1), Li W(2), Sumien N(2), Forster M(2), Simpkins JW(3), Liu R(2).
(1)Center for Neuroscience Discovery, University of North Texas Health Science
Center, Fort Worth, TX 76107, USA. Electronic address: [email protected].
(2)Center for Neuroscience Discovery, University of North Texas Health Science
Center, Fort Worth, TX 76107, USA. (3)Department of Physiology and Pharmacology,
Center for Neuroscience, Health Science Center, West Virginia University, Medical
Center Drive, Morgantown, WV 26506, USA.
Brain has exceptional high requirement for energy metabolism with glucose as the
exclusive energy source. Decrease of brain energy metabolism and glucose uptake
has been found in patients of Alzheimer's, Parkinson's and other
neurodegenerative diseases, providing a clear link between neurodegenerative
disorders and energy metabolism. On the other hand, cancers, including
glioblastoma, have increased glucose uptake and rely on aerobic glycolysis for
energy metabolism. The switch of high efficient oxidative phosphorylation to low
efficient aerobic glycolysis pathway (Warburg effect) provides macromolecule for
biosynthesis and proliferation. Current research indicates that methylene blue, a
century old drug, can receive electron from NADH in the presence of complex I and
donates it to cytochrome c, providing an alternative electron transfer pathway.
Methylene blue increases oxygen consumption, decrease glycolysis, and increases
glucose uptake in vitro. Methylene blue enhances glucose uptake and regional
cerebral blood flow in rats upon acute treatment. In addition, methylene blue
provides protective effect in neuron and astrocyte against various insults in
vitro and in rodent models of Alzheimer's, Parkinson's, and Huntington's disease.
In glioblastoma cells, methylene blue reverses Warburg effect by enhancing
mitochondrial oxidative phosphorylation, arrests glioma cell cycle at s-phase,
and inhibits glioma cell proliferation. Accordingly, methylene blue activates
AMP-activated protein kinase, inhibits downstream acetyl-coA carboxylase and
cyclin-dependent kinases. In summary, there is accumulating evidence providing a
proof of concept that enhancement of mitochondrial oxidative phosphorylation via
alternative mitochondrial electron transfer may offer protective action against
neurodegenerative diseases and inhibit cancers proliferation.
J Clin Endocrinol Metab. 2002 Nov;87(11):5325-31.
Progesterone production and actions in the human central nervous system and
neurogenic tumors.
Inoue T, Akahira J, Suzuki T, Darnel AD, Kaneko C, Takahashi K, Hatori M,
Shirane R, Kumabe T, Kurokawa Y, Satomi S, Sasano H.
Department of Pathology, Second Department of Surgery, Tohoku University School
of Medicine, 2-1 Seiryo-machi, Aoba-ku, Sendai 980-8575, Japan.
[email protected]
Progesterone has been suggested to be involved in the functions of the nervous
system, but it has yet to be examined in humans. Progesterone has also been
postulated to be involved in the biological behavior of various human neurogenic
tumors via progesterone receptors A and B (PR-A and PR-B). In this study we
examined the expression of PR and the enzymes responsible for progesterone
biosynthesis (P450scc, 3betahydroxysteroid dehydrogenase, and steroidogenic
acute regulatory protein) in human brain. We also examined the distribution of
PR isoforms in neurogenic tumors using immunohistochemistry and RT-PCR analysis.
The presence of PR and mRNA for P450scc, 3beta-hydroxysteroid dehydrogenase, and
steroidogenic acute regulatory protein was detected in human brain. PR isoforms
were detected in neurogenic tumors. PR-A and PR-B were equally expressed in
meningiomas, but PR-B was the predominant isoform compared with PR-A in
astrocytic tumors and Schwannomas. There was a statistically significant inverse
correlation between PR-A and the proliferation index in meningiomas and
astrocytic tumors. These findings suggest that progesterone is locally
synthesized and exerts its actions through PR in the human central nervous
system, and that progesterone may be involved in regulation of the growth and
development of neurogenic tumors via PR, especially in the inhibition of tumor
cell proliferation via PR-A.
Eur J Pharm Biopharm. 2016 Dec;109:81-92.
Inhibition of carbonic anhydrase IX in glioblastoma multiforme.
Amiri A(1), Le PU(2), Moquin A(1), Machkalyan G(1), Petrecca K(2), Gillard JW(3),
Yoganathan N(4), Maysinger D(5).
(1)Department of Pharmacology and Therapeutics, McGill University, Montreal,
Quebec H3G 1Y6, Canada.
(2)Department of Neurology and Neurosurgery, Montreal Neurological Institute and
Hospital, McGill University, Montreal H3A 2B4, Quebec, Canada.
(3)Department of Pharmacology and Therapeutics, McGill University, Montreal,
Quebec H3G 1Y6, Canada; Kalgene Pharmaceuticals, Innovation Park at Queens
University, Kingston K7L 3N6, Ontario, Canada.
(4)Kalgene Pharmaceuticals, Innovation Park at Queens University, Kingston K7L
3N6, Ontario, Canada.
(5)Department of Pharmacology and Therapeutics, McGill University, Montreal,
Quebec H3G 1Y6, Canada. Electronic address: [email protected].
Carbonic anhydrase IX (CAIX) is a transmembrane enzyme upregulated in several
types of tumors including glioblastoma multiforme (GBM). GBM is among the most
aggressive tumors among gliomas. Temozolomide (TMZ) therapy combined with
surgical or radiation approaches is the standard treatment but not effective in
long term. In this study we tested the treatment with acetazolamide (ATZ), an
inhibitor of CAIX, alone or combined with TMZ. The experiments were performed in
2D and 3D cultures (spheroids) using glioblastoma U251N and human brain tumor
stem cells (BTSCs). Several proteins implicated in tumor cell death were also
investigated. The key results from these studies suggest the following: (1) Cell
death of human glioblastoma spheroids and BTSC is significantly increased with
combined treatment after 7 days, and (2) the effectiveness of ATZ is
significantly enhanced against BTSC and U251N when incorporated into
nano-carriers. Collectively, these results point toward the usefulness of
nano-delivery of CAIX inhibitors and their combination with chemotherapeutics for
glioblastoma treatment.
Bioorg Med Chem. 2013 Jul 1;21(13):3949-57.
Hypoxia induced CA9 inhibitory targeting by two different sulfonamide derivatives
including acetazolamide in human glioblastoma.
Said HM(1), Hagemann C, Carta F, Katzer A, Polat B, Staab A, Scozzafava A,
Anacker J, Vince GH, Flentje M, Supuran CT.
(1)Dept. of Radiation Oncology, University of Würzburg, Germany.
[email protected]
HIF-1α regulated genes are mainly responsible for tumour resistance to radiation-
and chemo-therapy. Among these genes, carbonic anhydrase isoform IX (CA9) is
highly over expressed in many types of cancer especially in high grade brain
cancer like Glioblastoma (GBM). Inhibition of the enzymatic activity by
application of specific chemical CA9 inhibitor sulphonamides (CAI) like
Acetazolamide (Aza.), the new sulfonamide derivative carbonic anhydrase inhibitor
(SU.D2) or indirect inhibitors like the HIF-1α inhibitor Chetomin or molecular
inhibitors like CA9-siRNA are leading to an inhibition of the functional role of
CA9 during tumorigenesis. Human GBM cells were treated with in vitro hypoxia (1,
6, or 24 h at 0.1%, O2). Aza. application was at a range between 250 and 8000 nM
and the HIF-1α inhibitor Chetomin at a concentration range of 150-500 nM. Cell
culture plates were incubated for 24 h under hypoxia (0.1% O2). Further,
CA9-siRNA constructs were transiently transfected into GBM cells exposed to
extreme hypoxic aeration conditions. CA9 protein expression level was detectable
in a cell-type specific manner under normoxic conditions. Whereas U87-MG
exhibited a strong aerobic expression, U251 and U373 displayed moderate and GaMG
very weak normoxic CA9 protein bands. Aza. as well as SU.D2 displayed inhibitory
characteristics to hypoxia induced CA9 expression in the four GBM cell lines for
24 h of hypoxia (0.1% O2) at concentrations between 3500 and 8000 nM, on both the
protein and mRNA level. Parallel experiments using CA9-siRNA confirmed these
results. Application of 150-500 nM of the glycolysis inhibitor Chetomin under
similar oxygenation conditions led to a sharply reduced expression of both CA IX
protein and CA9 mRNA levels, indicating a clear glucose availability involvement
for the hypoxic HIF-1α and CA9 expression in GBM cells. Hypoxia significantly
influences the behaviour of human tumour cells by activation of genes involved in
the adaptation to hypoxic stress. The main objective in malignant GBM therapy is
either to eradicate the tumour or to convert it into a controlled, quiescent
chronic disease. Aza., SU.D2, Chetomin or CA9-siRNA possesses functional CA9
inhibitory characteristics when applied against human cancers with hypoxic
regions like GBM. They may be used as alternative or in conjunction with other
direct inhibitors possessing similar functionality, thereby rendering them as
potential optimal tools for the development of an optimized therapy in human
brain cancer treatment.
Neurosci Lett. 2004 Sep 30;368(3):279-84.
Aromatase, the enzyme responsible for estrogen biosynthesis, is expressed by
human and rat glioblastomas.
Yague JG(1), Lavaque E, Carretero J, Azcoitia I, Garcia-Segura LM.
(1)Instituto Cajal, C.S.I.C., Avenida Doctor Arce 37, E-28002 Madrid, Spain.
The biosynthesis of estradiol and related estrogens is catalyzed by the enzyme
aromatase. Among other tissues, aromatase is expressed in the brain, where it is
involved in the regulation of neuroendocrine events and reproduction. Under
physiological conditions, the expression of aromatase in the mammalian brain is
restricted to neurons. However, recent studies have shown that reactive
astrocytes express aromatase after brain injury. This opens the possibility for
the expression of the enzyme in other altered forms of glial cell, such as
gliomas. In the present study, the expression of aromatase has been assessed, by
RT-PCR and immunocytochemistry, in the rat glioblastoma C6 and in two human
glioblastoma cell lines T98G and U373MG. The three cell lines expressed aromatase
mRNA and showed a cytoplasmic pattern of aromatase immunoreactivity. In addition,
the three cell lines express estrogen receptor alpha, suggesting that estradiol
formed by aromatase may act as an autocrine or paracrine factor for glioblastoma
cells. By analogy to the implication of aromatase into the growth of other forms
of estrogen-sensitive tumors, such as some breast cancers, it is conceivable that
the expression of aromatase may play a role in the growth of glioblastomas.
Oncotarget. 2017 Mar 7;8(10):16605-16620.
Anti-GD2-ch14.18/CHO coated nanoparticles mediate glioblastoma (GBM)-specific
delivery of the aromatase inhibitor, Letrozole, reducing proliferation, migration
and chemoresistance in patient-derived GBM tumor cells.
Tivnan A(1), Heilinger T(1)(2), Ramsey JM(3), O'Connor G(3), Pokorny JL(4)(5),
Sarkaria JN(4), Stringer BW(6), Day BW(6), Boyd AW(6), Kim EL(7), Lode HN(8),
Cryan SA(3), Prehn JH(1).
(1)Centre for Systems Medicine, Department of Physiology and Medical Physics,
Royal College of Surgeons in Ireland, York House, Dublin 2, Ireland.
(2)IMC Fachhochschule Krems, University of Applied Sciences, Krems, Austria.
(3)School of Pharmacy, Royal College of Surgeons in Ireland, York House, Dublin
2, Ireland & Tissue Engineering Research Group, Department of Anatomy, RCSI and
Centre for Research in Medical Devices (CURAM), NUIG, Ireland.
(4)Department of Radiation Oncology, Mayo Clinic, Rochester, MN, United States of
America.
(5)Department of Neurosurgery, Stanford University, Stanford, CA 94305, USA.
(6)Brain Cancer Research Unit, QIMR Berghofer Medical Research Institute,
Brisbane, Australia.
(7)Laboratory of Neurooncology, Department of Neurosurgery, Johannes Gutenberg
University Medical Center, Mainz, Germany.
(8)Department of Paediatrics and Paediatric Haematology/Oncology, University of
Greifswald, Greifswald, Germany.
Aromatase is a critical enzyme in the irreversible conversion of androgens to
oestrogens, with inhibition used clinically in hormone-dependent malignancies. We
tested the hypothesis that targeted aromatase inhibition in an aggressive brain
cancer called glioblastoma (GBM) may represent a new treatment strategy. In this
study, aromatase inhibition was achieved using third generation inhibitor,
Letrozole, encapsulated within the core of biodegradable poly lactic-co-glycolic
acid (PLGA) nanoparticles (NPs). PLGA-NPs were conjugated to human/mouse chimeric
anti-GD2 antibody ch14.18/CHO, enabling specific targeting of GD2-positive GBM
cells. Treatment of primary and recurrent patient-derived GBM cells with
free-Letrozole (0.1 μM) led to significant decrease in cell proliferation and
migration; in addition to reduced spheroid formation. Anti-GD2-ch14.18/CHO-NPs
displayed specific targeting of GBM cells in colorectal-glioblastoma co-culture,
with subsequent reduction in GBM cell numbers when treated with
anti-GD2-ch14.18-PLGA-Let-NPs in combination with temozolomide. As miR-191 is an
estrogen responsive microRNA, its expression, fluctuation and role in Letrozole
treated GBM cells was evaluated, where treatment with premiR-191 was capable of
rescuing the reduced proliferative phenotype induced by aromatase inhibitor. The
repurposing and targeted delivery of Letrozole for the treatment of GBM, with the
potential role of miR-191 identified, provides novel avenues for target
assessment in this aggressive brain cancer.
J Neurooncol. 2014 Sep;119(2):275-84.
Aromatase and estrogen receptor alpha mRNA expression as prognostic biomarkers in
patients with astrocytomas.
Dueñas Jiménez JM(1), Candanedo Arellano A, Santerre A, Orozco Suárez S, Sandoval
Sánchez H, Feria Romero I, López-Elizalde R, Alonso Venegas M, Netel B, de la
Torre Valdovinos B, Dueñas Jiménez SH.
(1)Centro Universitario de Ciencias de la Salud, Universidad de Guadalajara,
Guadalajara, Jalisco, Mexico, [email protected].
Estrogens are oncogenic hormones at a high level in breast, prostate, endometrial
and lung cancer. Estrogens are synthesized by aromatase which has been used as a
biomarker both in breast and lung cancer. Estrogen biological activities are
executed by their classic receptors (ERα and ERβ). ERα has been described as a
cancer promoter and ERβ, as a possible tumor suppressor. Both receptors are
present at low levels in primary multiforme glioblastoma (GBM). The GBM frequency
is 50 % higher in men than in women. The GBM patient survival period ranges from
7 to 18 months. The purpose of this pilot study was to evaluate aromatase and
estrogen receptor expression, as well as 17ß-estradiol concentration in
astrocytoma patients biopsies to obtain a prognosis biomarker for these patients.
We analyzed 36 biopsies of astrocytoma patients with a different grade (I-IV) of
malignity. Aromatase and estrogen receptor mRNA expression were analyzed by
semiquantitative RT-PCR, and the E2 levels, by ELISA. E2 concentration was higher
in GBM, compared to grade II or III astrocytomas. The number of cells
immunoreactive to aromatase and estrogen receptors decreased as the grade of
tumor malignity increased. Aromatase mRNA expression was present in all biopsies,
regardless of malignity grade or patient age or gender. The highest expression of
aromatase mRNA in GBM patients was associated to the worst survival prognostic
(6.28 months). In contrast lowest expression of ERα mRNA in astrocytoma patients
had a worst prognosis. In conclusion, aromatase and ERα expression could be used
as prognosis biomarkers for astrocytoma patients.
Bioorg Med Chem. 2013 Jul 1;21(13):3949-57.
Hypoxia induced CA9 inhibitory targeting by two different sulfonamide derivatives
including acetazolamide in human glioblastoma.
Said HM(1), Hagemann C, Carta F, Katzer A, Polat B, Staab A, Scozzafava A,
Anacker J, Vince GH, Flentje M, Supuran CT.
(1)Dept. of Radiation Oncology, University of Würzburg, Germany.
[email protected]
HIF-1α regulated genes are mainly responsible for tumour resistance to radiation-
and chemo-therapy. Among these genes, carbonic anhydrase isoform IX (CA9) is
highly over expressed in many types of cancer especially in high grade brain
cancer like Glioblastoma (GBM). Inhibition of the enzymatic activity by
application of specific chemical CA9 inhibitor sulphonamides (CAI) like
Acetazolamide (Aza.), the new sulfonamide derivative carbonic anhydrase inhibitor
(SU.D2) or indirect inhibitors like the HIF-1α inhibitor Chetomin or molecular
inhibitors like CA9-siRNA are leading to an inhibition of the functional role of
CA9 during tumorigenesis. Human GBM cells were treated with in vitro hypoxia (1,
6, or 24 h at 0.1%, O2). Aza. application was at a range between 250 and 8000 nM
and the HIF-1α inhibitor Chetomin at a concentration range of 150-500 nM. Cell
culture plates were incubated for 24 h under hypoxia (0.1% O2). Further,
CA9-siRNA constructs were transiently transfected into GBM cells exposed to
extreme hypoxic aeration conditions. CA9 protein expression level was detectable
in a cell-type specific manner under normoxic conditions. Whereas U87-MG
exhibited a strong aerobic expression, U251 and U373 displayed moderate and GaMG
very weak normoxic CA9 protein bands. Aza. as well as SU.D2 displayed inhibitory
characteristics to hypoxia induced CA9 expression in the four GBM cell lines for
24 h of hypoxia (0.1% O2) at concentrations between 3500 and 8000 nM, on both the
protein and mRNA level. Parallel experiments using CA9-siRNA confirmed these
results. Application of 150-500 nM of the glycolysis inhibitor Chetomin under
similar oxygenation conditions led to a sharply reduced expression of both CA IX
protein and CA9 mRNA levels, indicating a clear glucose availability involvement
for the hypoxic HIF-1α and CA9 expression in GBM cells. Hypoxia significantly
influences the behaviour of human tumour cells by activation of genes involved in
the adaptation to hypoxic stress. The main objective in malignant GBM therapy is
either to eradicate the tumour or to convert it into a controlled, quiescent
chronic disease. Aza., SU.D2, Chetomin or CA9-siRNA possesses functional CA9
inhibitory characteristics when applied against human cancers with hypoxic
regions like GBM. They may be used as alternative or in conjunction with other
direct inhibitors possessing similar functionality, thereby rendering them as
potential optimal tools for the development of an optimized therapy in human
brain cancer treatment.

1. Tumour Biol. 2015 May;36(5):3407-15.
Fisetin suppresses ADAM9 expression and inhibits invasion of glioma cancer cells
through increased phosphorylation of ERK1/2.
Chen CM(1), Hsieh YH, Hwang JM, Jan HJ, Hsieh SC, Lin SH, Lai CY.
(1)Division of Neurosurgery, Department of Surgery, Changhua Christian Hospital,
Changhua, Taiwan.
Fisetin (3,3',4',7-tetrahydroxyflavone) is a naturally occurring flavonoid which
is widely distributed in plants. It has been reported to possess some anticancer
and anti-invasive capabilities. We set out to explore the effects of fisetin on
antimetastatic and its mechanism of action in GBM8401 cells. The results
indicated that fisetin exhibited effective inhibition of cell migration and
inhibited the invasion of GBM8401 cells under non-cytotoxic concentrations. To
identify the potential targets of fisetin, human proteinase antibody array
analysis was performed, and the results indicated that the fisetin treatment
inhibited the expression of ADAM9 protein and mRNA, which are known to contribute
to the progression of glioma cancer. Our results showed that fisetin
phosphorylated ERK1/2 in a sustained way that contributed to the inhibited ADAM9
protein and mRNA expression determined by Western blot and RT-PCR. Moreover,
inhibition of ERK1/2 by U0126 or transfection with the siERK plasmid
significantly abolished the fisetin-inhibited migration and invasion through
activation of the ERK1/2 pathway. In summary, our results suggest that fisetin
might be a potential therapeutic agent against human glioma cells based on its
capacity to activate ERK1/2 and to inhibit ADAM9 expression.
2. Nutr Cancer. 2016 Nov-Dec;68(8):1357-1368.
Inhibitory Kinetics and Mechanism of Flavonoids Extracted from Cotinus coggygria
Scop. Against Glioblastoma Cancer.
Wang G(1), Wang JJ(1)(2), Du L(1), Fei L(1), To ST(3). (1)a Department of Pharmaceutics , Shanghai Eighth People's Hospital, Jiangsu
University , Shanghai , China.
(2)b Hubei University of Medicine , Shiyan , Hubei , China.
(3)c Department of Health Technology and Informatics , The Hong Kong Polytechnic
University , Hong Kong.
This proposal seeks to study the potential therapeutic modality of
chemoprevention and anticancer effects and mechanisms of the flavonoids from
Cotinus coggygria Scop. on glioblastoma cancer. In the current study, the total
flavonoids (TFs) isolated from Cotinus coggygria Scop. var. cinerea Engl.
(Cotinus coggygria Scop.) and the major flavonoids of Cotinus coggygria Scop.
(CCFs) were identified, and the inhibitory kinetics of TF and CCF on glioblastoma
cell lines were calculated. We also investigated whether TF or CCF regulated the
apoptotic mechanism in cellular models of glio-blastoma cells. Finally, we
evaluated whether treatment with TF or CCF suppressed tumor growth and inhibited
migration in orthotopic mouse models of glioblastoma in vivo. In this study, the
CCFs were identified as rutin, myricetin, and fisetin. TF and CCF remarkably
inhibited cell proliferation and downregulated the PI3K/Akt and ERK signaling
pathway in glioblastoma cell lines. Furthermore, the mitochondrial
caspase-dependent cascade was regulated by TF and myricetin. In addition, TF and
myricetin exhibited significant antitumor effects on glioblastoma in vivo. Taken
together, these results suggest that phytochemical and biological data provide
evidence for the active components in Cotinus coggygria, and that the TFs are
responsible for the anticancer effects on glioblastoma cell growth via induction
of apoptosis. In addition, the representative compound myricetin could provide a
clinically relevant therapeutic opportunity. Therefore, our data strongly suggest
that myricetin-deprived CCF can serve as a potent chemopreventive herbal
medicine.
Biochem Biophys Res Commun. 2006 Apr 21;342(4):1329-33.
The effect of gamma-tocopherol on proliferation, integrin expression, adhesion, and migration of human glioma cells.
Samandari E, Visarius T, Zingg JM, Azzi A.
The effect of vitamin E on proliferation, integrin expression, adhesion, and migration in human glioma cells has been studied. gamma-tocopherol at 50 microM concentration exerted more inhibitory effect than alpha-tocopherol at the same concentration on glioma cell proliferation. Integrin alpha5 and beta1 protein levels were increased upon both alpha- and gamma-tocopherol treatments. In parallel, an increase in the alpha5beta1 heterodimer cell surface expression was observed. The tocopherols inhibited glioma cell-binding to fibronectin where gamma-tocopherol treatment induced glioma cell migration. Taken together, the data reported here are consistent with the notion that the inhibition of glioma cell proliferation induced by tocopherols may be mediated, at least in part, by an increase in integrin alpha5 and beta1 expression. Cell adhesion is also negatively affected by tocopherols, despite a small increase in the surface appearance of the alpha5beta1 heterodimer. Cell migration is stimulated by gamma-tocopherol. It is concluded that alpha5 and beta1 integrin expression and surface appearance are not sufficient to explain all the observations and that other integrins or in general other factors may be associated with these events.
Free Radic Biol Med. 2006 Aug 1;41(3):464-72.
Antiproliferative effects of tocopherols (vitamin E) on murine glioma C6 cells:
homologue-specific control of PKC/ERK and cyclin signaling.
Betti M(1), Minelli A, Canonico B, Castaldo P, Magi S, Aisa MC, Piroddi M, Di
Tomaso V, Galli F.
(1)Institute of Physiological Sciences, University of Urbino Carlo Bo, Urbino,
Italy.
Chemoprevention strategies for brain tumors (specifically gliomas) are few and
surprisingly poorly investigated. We have studied the effects of tocopherols
(TOCs; vitamin E) on proliferation and death processes of murine glioma C6 cells.
These vitamers showed different cell uptake and concentration- and time-dependent
inhibitory effects on cell growth that were significant at the lowest
concentrations tested (1-10 microM). However, the inhibitory potency of TOCs
seemed to reflect at least in part their actual cell concentrations at steady
state, with the order of magnitude gamma-TOC >or= alpha-TOC > delta-TOC
approximately or = beta-TOC. Moreover, for extracellular concentrations >or=10
microM, TOCs also showed a significant cytotoxic effects due mainly to necrosis,
while apoptosis was negligible. Gamma-TOC (the form showing preferential cell
uptake and lowest unspecific cytotoxicity) was the most effective inhibitor of
cell cycle progression (arrest in G0/G1 phase) leading to lowered expression of
cyclin E and cyclin-dependent kinases 2 and 4 and overexpression of p27 (specific
inhibitor of S-phase entering). According to these signals, activated ERK1/2 and
PKC upstream and Rb phosphorylation downstream were decreased. In conclusion,
within TOCs the gamma form exerts the most potent and specific control of cell
cycle progression in C6 cells (cytostatic effect). This suggests a
chemopreventive role of this form of vitamin E in gliomas.
Proc Soc Exp Biol Med. 1983 Nov;174(2):302-7.
Study on the specificity of alpha-tocopheryl (vitamin E) acid succinate effects
on melanoma, glioma and neuroblastoma cells in culture.
Rama BN, Prasad KN.
d- and dl-alpha-tocopheryl succinate inhibited growth and caused morphological
changes in mouse melanoma (B-16), mouse neuroblastoma (NBP2), and rat glioma
(C-6) cells in culture. To study whether the effects of alpha-tocopheryl (vitamin
E) succinate on tumor cells are mediated by antioxidant mechanisms, the effects
of lipid-soluble antioxidants, butylated hydroxyanisole (BHA) and butylated
hydroxytoluene (BHT) were compared with those of vitamin E succinate. Results
showed that these antioxidants produced alterations on the growth and morphology
of neuroblastoma, melanoma, and glioma cells which are similar to those produced
by vitamin E succinate; however, the extent of the effect depended upon the type
of antioxidant and the form of tumor cells. These data suggest that the effects
of vitamin E succinate on tumor cells may be mediated, in part, by antioxidant
mechanisms.
Neurol Med Chir (Tokyo). 1983 Jun;23(6):421-7.
[Effect of vitamin E (DL-alpha-tocopherol) on the growth of the rat glioma cells
in culture].
[Article in Japanese]
Kokunai T, Korosue K, Tamaki N, Matsumoto S.

No Shinkei Geka. 1999 Feb;27(2):119-25.
[Proliferation inhibition of glioma cells by vitamin K2].
[Article in Japanese]
Sun L(1), Yoshii Y, Miyagi K, Ishida A.
(1)Department of Neurosurgery, School of Medicine, University of the Ryukyus,
Okinawa, Japan.
The antitumor effects of vitamin K2 were studied using three glioma cell lines:
C6 (rat glioma cell), RBR17T and T98G (human glioma cell). The antitumor effects
were estimated by count assay. The results was that vitamin K2 induced growth
inhibition in a dose-dependent manner. The RBR 17T cells exposed to vitamin K2
for 72 hours resulted in oligonucleosomal DNA fragmentation and formed a ladder
on agarose gel electrophoresis. Furthermore, the RBR17T cells exposed to vitamin
K2 for 24 hours were significantly accumulated in the G0G1 phase of the cell
cycle. Those results suggested that vitamin K2 can inhibit the proliferation of
cells through the induction of cell cycle arrest and apoptosis for tumor cells.
The combined treatment of vitamin K2 with ACNU or 5-FU or INF-beta or
1,25-dihydroxyvitamin D3 enhanced growth inhibition significantly. In conclusion,
vitamin K2 can be a useful drug for the treatment of glioma.

Acta Neurochir (Wien). 1995;133(3-4):184-90.
Effects of vitamin D and retinoic acid on human glioblastoma cell lines.
Magrassi L(1), Butti G, Pezzotta S, Infuso L, Milanesi G.
(1)Department of Surgery, University of Pavia, IRCCS Policlinico S. Mattco,
Italy.
The biological significance of vitamin D receptors expressed by glioblastoma and
other glial tumours is still unclear. In an effort to clarify this issue we
studied the effects of increasing concentrations of 25-dihydroxyvitamin D3 and
its metabolite 1 alpha,25-dihydroxyvitamin D3 on two human glioblastoma cell
lines. Both substances were capable of inducing a significant (> 50%) reduction
in growth of the two glioblastoma cell lines at dosages over 5 microM. When the
HU 70 cell line was treated by increasing dilutions of 25-dihydroxyvitamin D3
combined with 1 microM all trans-retinoic acid, significant inhibition was
apparent even after addition of 25-dihydroxyvitamin D3 in the nanomolar range.
Reduction of growth index was mainly due to induced cell death. Our results
provide in vitro evidence that vitamin D metabolites alone or in combination with
retinoids may be potentially useful agents in the differentiation therapy of
human malignant gliomas.

 

[goodandevil]

On Naltrexone dosage to stimulate lutenizing hormone:

Q: "I have 50mg pills, do you think the low-dose or normal dose is best, and for how long to stimulate LH would you say? "

R: "My experience has been with doses of one or two milligrams, repeated for two or three days, when the effect is achieved, then waiting two weeks or more to see if repeating the process is needed."

 

[Wagner83]

Me:
Hi Ray,

I've looked a bit into minerals, vitamins and their balance which affect enzymes which in turn affect hormones. So in this regard I wonder what would be the purpose to manipulate hormones rather than minerals/vitamins intake, and last but not least, how would supplementing even low doses of hormones not lead to minerals/vitamins imbalances?
From what I understand enzymes are tightly regulated by nutrients, and if not enough of the right nutrients are provided then the body will not function properly, similarly, supplementing hormones may resolve issues in the short term but then induce deficiencies etc..

Ray:
The body has a powerful selective ability, able to extract things against a concentration gradient. For example, when hair is stripped of minerals and then dipped into a solution of minerals similar to the serum (high sodium, low potassium, etc.) the hair will take up potassium in preference to sodium. And the hair is dead—living cells have even greater selectivity. It’s energy that maintains the structural selectivity; thyroid provides energy, cholesterol/pregnenolone/
progesterone/DHEA reinforce structure.

Me:

Thanks, do you have any references I can look into regarding this topic of
Energy maintaining structure and its selectivity?

What would be the negative consequences of normalizing stress hormones through direct intervention? Without meeting the condition that got them to become elevated in the first place.


Raymond:

When cortisol is increased, though it does reduce inflammation and reduce leakiness, it’s antagonistic to androgens, and shifts the balance toward protein catabolism, and lowers the oxidation of glucose.

Related to selectivity of energized cytoplasm---D.N. Nasonov, V.I. Vernadsky, H.G. Bungenberg de Jong, G.N. Ling, A.S. Troshin, V. Matveev, Sidney Fox, and recently I think Jeremy L. England is working on related things, energy creating order.

Me:

If serotonin or estrogen were involved rather than cortisol, would you have the same approach?

Do you know if particular conditions could lead to high cortisol, serotonin or estrogen and unless they are addressed the need for supplemental hormones or drugs will always be present?
The only people I've seen use hormonal supplements and not need them anymore used thyroid, but most people need the supplements continuously and quite a few had troubles with dhea, pregnenolone or thyroid. Micro-dosing psilocybin or lsd appears to benefit all and relieve depression but again the benefits vanish upon cessation.

Ray:

Although occasionally a short supplementation of progesterone, pregnenolone, or thyroid will decisively correct a problem, most often there has been something environmental-nutritional that had to be changed. If people don’t change their diet and/or problematic surroundings, then they are dependent on hormones, stimulants, antiinflammatories, etc.

 

[charlie]

Reminder!

This thread is intended as a depository of email's from Ray Peat. This thread is not for discussions of those emails. If you would like to discuss contents of an email simply make another thread or post in the "Discussion Thread".

 

[Miso]

Q:I would like to know your take on the Hydrogen Gas as a therapy, with more recent studies in mice showing pretty significant protective effects. Is hydrogen gas able to be absorb into the bloodstream and diffuse into the cells as an antioxidant, and would it differ from normal hydrogen gas produced by the gut bacteria? Perhaps concentration? There are some hydrogen gas producing generators base on electrolysis, and there are elemental magnesium based tablets that can be added to water. Are these worth testing?

A: I would be cautious with the hydrogen generating tablets, but hydrogen gas is plausible as an antiinflammatory supplement. I think any produced by bacteria might be beneficial too.

Q: Do you think the Hydrogen producing tablets risk is associated with alkalinity/Hydroxide formation or heavy metals?

A: Impurities would be my concern with the tablets.

Methods Enzymol. 2015;555:289-317.
Molecular hydrogen as a novel antioxidant: overview of the advantages of hydrogen
for medical applications.
Ohta S(1).
(1)Department of Biochemistry and Cell Biology, Institute of Development and
Aging Sciences, Graduate School of Medicine, Nippon Medical School, Kawasaki,
Japan. Electronic address: [email protected].
Molecular hydrogen (H2) was believed to be inert and nonfunctional in mammalian
cells. We overturned this concept by demonstrating that H2 reacts with highly
reactive oxidants such as hydroxyl radical ((•)OH) and peroxynitrite (ONOO(-))
inside cells. H2 has several advantages exhibiting marked effects for medical
applications: it is mild enough neither to disturb metabolic redox reactions nor
to affect signaling by reactive oxygen species. Therefore, it should have no or
little adverse effects. H2 can be monitored with an H2-specific electrode or by
gas chromatography. H2 rapidly diffuses into tissues and cells to exhibit
efficient effects. Thus, we proposed the potential of H2 for preventive and
therapeutic applications. There are several methods to ingest or consume H2:
inhaling H2 gas, drinking H2-dissolved water (H2-water), injecting H2-dissolved
saline (H2-saline), taking an H2 bath, or dropping H2-saline onto the eyes.
Recent publications revealed that, in addition to the direct neutralization of
highly reactive oxidants, H2 indirectly reduces oxidative stress by regulating
the expression of various genes. Moreover, by regulating gene expression, H2
functions as an anti-inflammatory, antiallergic, and antiapoptotic molecule, and
stimulates energy metabolism. In addition to growing evidence obtained by model
animal experiments, extensive clinical examinations were performed or are under
way. Since most drugs specifically act on their specific targets, H2 seems to
differ from conventional pharmaceutical drugs. Owing to its great efficacy and
lack of adverse effects, H2 has potential for clinical applications for many
diseases.

Surgery. 2015 Aug;158(2):399-407.
Hydrogen inhalation protects against acute lung injury induced by hemorrhagic
shock and resuscitation.
Kohama K(1), Yamashita H(2), Aoyama-Ishikawa M(2), Takahashi T(3), Billiar TR(4),
Nishimura T(1), Kotani J(1), Nakao A(5).
(1)Department of Emergency, Disaster and Critical Care Medicine, Hyogo College of
Medicine, Nishinomiya, Japan.
(2)Kobe University Graduate School of Health Science, Kobe, Hyogo, Japan.
(3)Faculty of Health and Welfare Science, Okayama Prefectural University,
Okayama, Japan.
(4)Department of Surgery, University of Pittsburgh, Pittsburgh, PA.
(5)Department of Emergency, Disaster and Critical Care Medicine, Hyogo College of
Medicine, Nishinomiya, Japan. Electronic address: [email protected].
INTRODUCTION: Hemorrhagic shock followed by fluid resuscitation (HS/R) triggers
an inflammatory response and causes pulmonary inflammation that can lead to acute
lung injury (ALI). Hydrogen, a therapeutic gas, has potent cytoprotective,
antiinflammatory, and antioxidant effects. This study examined the effects of
inhaled hydrogen on ALI caused by HS/R.
METHODS: Rats were subjected to hemorrhagic shock by withdrawing blood to lower
blood pressure followed by resuscitation with shed blood and saline to restore
blood pressure. After HS/R, the rats were maintained in a control gas of similar
composition to room air or exposed to 1.3% hydrogen.
RESULTS: HS/R induced ALI, as demonstrated by significantly impaired gas
exchange, congestion, edema, cellular infiltration, and hemorrhage in the lungs.
Hydrogen inhalation mitigated lung injury after HS/R, as indicated by
significantly improved gas exchange and reduced cellular infiltration and
hemorrhage. Hydrogen inhalation did not affect hemodynamic status during HS/R.
Exposure to 1.3% hydrogen significantly attenuated the upregulation of the
messenger RNAs for several proinflammatory mediators induced by HS/R. Lipid
peroxidation was reduced significantly in the presence of hydrogen, indicating
antioxidant effects.
CONCLUSION: Hydrogen, administered through inhalation, may exert potent
therapeutic effects against ALI induced by HS/R and attenuate the activation of
inflammatory cascades.
Copyright © 2015 Elsevier Inc. All rights reserved.

 

[member 2106]

I asked Ray about ways to reduce swelling of the prostate.

Ray said: "Inflammation in the intestine can raise systemic or regional histamine and serotinin enough to cause prostate swelling. Avoiding starchy vegetables, might help; supplementing vitamin D and thyroid can reduce inflammation. An antihistamine (diphenhydrame or cyproheptadine) and aspirin can reduce prostate swelling, and might help the intestine too."