Pyruvate carboxylase (PC) is a mitochondrial, biotin-containing enzyme catalyzing the ATP-dependent synthesis of oxaloacetate from pyruvate and bicarbonate, with a critical anaplerotic role in sustaining the brain metabolism. Based on the studies performed on animal models, PC expression was assigned to be glia-specific. To study PC distribution among human neural cells, we probed the cultured human astrocytes and brain sections with antibodies against PC. Additionally, we tested the importance of PC for the viability of cultured human astrocytes by applying the PC inhibitor 3-chloropropane-1,2-diol (CPD). Our results establish the expression of PC in mitochondria of human astrocytes in culture and brain tissue and also into a subpopulation of the neurons in situ. CPD negatively affected the viability of astrocytes in culture, which could be partially reversed by supplementing media with malate, 2-oxoglutarate, citrate, or pyruvate. The provided data estimates PC expression in human astrocytes and neurons in human brain parenchyma. Furthermore, the enzymatic activity of PC is vital for sustaining the viability of cultured astrocytes.
- MeSH
- astrocyty * metabolismus MeSH
- kyselina pyrohroznová metabolismus MeSH
- lidé MeSH
- mozek metabolismus MeSH
- neurony metabolismus MeSH
- pyruvátkarboxylasa * metabolismus MeSH
- zvířata MeSH
- Check Tag
- lidé MeSH
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
The role of immune system in carcinogenesis represents fundamental events associated with cancer eradication; however, tumor evolution is connected with various mechanisms of tumor evasion and progression of cancer. Based on recent evidence, phytochemicals are directly associated with immunomodulation of the innate and adaptive immunity via different mechanisms of action including stimulation and amplification of immune cells, humoral compartments, and associated molecules. This comprehensive study focuses on immunomodulating potential of phytochemicals (mixture in plants or separately such as individual phytochemical) and their impact on regulation of immune response during cancer development, immune tolerance, and immune escape. Clinical application of phytochemicals as modulators of host immunity against cancer may represent perspective approach in anticancer therapy.
Cieľ: Včasné rozpoznanie sclerosis multiplex (SM) pomáha začať liečbu pacientov skôr, a tak oddialiť progresiu ochorenia. Urobili sme analýzu metabolitov cerebro-spinálneho likvoru (cerebrospinal fluid; CSF), s cieľom zistiť prediktory včas, a tak oddialiť SM. Metódy: Do štúdie bolo zaradených 56 jedincov s podozrením na SM, pred začatím akejkoľvek liečby. Z nich bolo 28 diagnostikovaných ako definitívna SM, u 17 pacientov sme zistili klinicky izolovaný syndróm (clinically isolated syndrome; CIS) podľa McDonaldových kritérií z roku 2010, v 11 prípadoch sa jednalo o iné demyelinizačné ochorenie CNS (DEM). Kontrolnú skupinu (CON) tvorili 29 jedinci, ktorí nemali dokázané žiadne ochorenie CNS. Na meranie metabolitov CSF bola použitá protonová nukleárna magnetická rezonančná spektroskopia. Výsledky: Glutamín, ktorý koreloval s Expanded Disability Status Scale (EDSS), bol jediným metabolitom, ktorý dokázal odlíšiť CIS, SM, DEM a CON. Valín, leucín, isoleucín, znížené u CIS a SM v porovnaní s CON, sa neodlišovali od DEM. Hladiny citrátu v CSF špecifikovali SM a CIS oproti DEM, ale nepomohli v rozlíšení CIS a SM. Citrát ukazoval signifikantné korelácie s vekom, dľžkou trvania ochorenia a EDSS u SM pacientov. Acetát, aceton, pyruvát, formát, histidin v CSF neboli signifikantnými prediktormi SM alebo CIS, hoci korelovali s niektorými vybranými premennými. Záver: Táto práca ukazuje prediktívnu úlohu glutamínu v CSF v stanovení diagnózy SM od jej včasných štádií, vypichujúc tak dôležitú úlohu glutamát/glutamínového cyklu v patogenéze SM. Ďalší potenciálny prediktor SM bol citrát. Ďalšie metabolity neboli identifikované ako senzitívne CSF markery SM.
Aim: Early recognition of multiple sclerosis (MS) allows patients to begin treatment earlier and delay disease progression. We performed an analysis of cerebrospinal fluid (CSF) metabolites to find early predictors of MS. Methods: We included 56 participants with suspected MS before any treatment. Out of those, 28 patients were diagnosed with definite MS, 17 with clinically isolated syndrome (CIS) according to McDonald 2010 criteria, and 11 with other demyelinating diseases (DEM) of the CNS. The control group (CON) included 29 participants without any confirmed CNS disease. Proton nuclear magnetic resonance spectroscopy was used to measure CSF metabolites. Results: Glutamine, correlating with Expanded Disability Status Scale (EDSS), was the only metabolite capable to distinguish between CIS and MS, DEM, and CON. Valine, leucine, isoleucine, decreased in CIS and MS when compared with CON, did not differ from DEM. Citrate CSF levels specified MS and CIS against DEM but did not help to distinguish between CIS and MS. Citrate showed significant correlations with age, disease duration, and EDSS in MS patients. Acetate, acetone, pyruvate, formate and histidine CSF levels were not significant predictors of MS or CIS, although they correlated with selective variables. Conclusion: This work shows the predictive role of CSF glutamine in diagnosing MS since its early stages, pinpointing an important role of the glutamate/glutamine cycle in MS pathogenesis. Another potential predictor of MS was citrate. Other metabolites were not identified as sensitive CSF markers of MS.
- MeSH
- demyelinizační nemoci diagnostické zobrazování MeSH
- klinická studie jako téma MeSH
- lidé MeSH
- magnetická rezonanční spektroskopie metody MeSH
- metabolomika MeSH
- mozkomíšní mok diagnostické zobrazování MeSH
- roztroušená skleróza * diagnóza MeSH
- Check Tag
- lidé MeSH
- Publikační typ
- práce podpořená grantem MeSH
Comprehensive oncology research suggests an important role of phytochemicals or whole plant foods in the modulation of signaling pathways associated with anticancer action. The goal of this study is to assess the anticancer activities of Cinnamomum zeylanicum L. using rat, mouse, and cell line breast carcinoma models. C. zeylanicum (as bark powder) was administered in the diet at two concentrations of 0.1% (w/w) and 1% (w/w) during the whole experiment in chemically induced rat mammary carcinomas and a syngeneic 4T1 mouse model. After autopsy, histopathological and molecular evaluations of mammary gland tumors in rodents were carried out. Moreover, in vitro analyses using MCF-7 and MDA-MB-231 cells were performed. The dominant metabolites present in the tested C. zeylanicum essential oil (with relative content over 1%) were cinnamaldehyde, cinnamaldehyde dimethyl acetal, cinnamyl acetate, eugenol, linalool, eucalyptol, limonene, o-cymol, and α-terpineol. The natural mixture of mentioned molecules demonstrated significant anticancer effects in our study. In the mouse model, C. zeylanicum at a higher dose (1%) significantly decreased tumor volume by 44% when compared to controls. In addition, treated tumors showed a significant dose-dependent decrease in mitotic activity index by 29% (0.1%) and 45.5% (1%) in comparison with the control group. In rats, C. zeylanicum in both doses significantly reduced the tumor incidence by 15.5% and non-significantly suppressed tumor frequency by more than 30% when compared to controls. An evaluation of the mechanism of anticancer action using valid oncological markers showed several positive changes after treatment with C. zeylanicum. Histopathological analysis of treated rat tumor specimens showed a significant decrease in the ratio of high-/low-grade carcinomas compared to controls. In treated rat carcinomas, we found caspase-3 and Bax expression increase. On the other hand, we observed a decrease in Bcl-2, Ki67, VEGF, and CD24 expressions and MDA levels. Assessment of epigenetic changes in rat tumor cells in vivo showed a significant decrease in lysine methylation status of H3K4m3 and H3K9m3 in the high-dose treated group, a dose-dependent increase in H4K16ac levels (H4K20m3 was not changed), down-regulations of miR21 and miR155 in low-dose cinnamon groups (miR22 and miR34a were not modulated), and significant reduction of the methylation status of two out of five gene promoters-ATM and TIMP3 (PITX2, RASSF1, PTEN promoters were not changed). In vitro study confirmed results of animal studies, in that the essential oil of C. zeylanicum displayed significant anticancer efficacy in MCF-7 and MDA-MB-231 cells (using MTS, BrdU, cell cycle, annexin V/PI, caspase-3/7, Bcl-2, PARP, and mitochondrial membrane potential analyses). As a conclusion, C. zeylanicum L. showed chemopreventive and therapeutic activities in animal breast carcinoma models that were also significantly confirmed by mechanistic evaluations in vitro and in vivo.
- MeSH
- antitumorózní látky fytogenní aplikace a dávkování chemie farmakologie MeSH
- histony metabolismus MeSH
- krysa rodu rattus MeSH
- kůra rostlin chemie MeSH
- lidé MeSH
- MFC-7 buňky MeSH
- mikro RNA genetika MeSH
- myši MeSH
- nádorové buněčné linie MeSH
- nádory prsu farmakoterapie genetika metabolismus MeSH
- oleje prchavé aplikace a dávkování chemie farmakologie MeSH
- oleje rostlin aplikace a dávkování chemie farmakologie MeSH
- proliferace buněk účinky léků MeSH
- skořicovník ceylonský chemie MeSH
- viabilita buněk účinky léků MeSH
- vztah mezi dávkou a účinkem léčiva MeSH
- xenogenní modely - testy antitumorózní aktivity MeSH
- zvířata MeSH
- Check Tag
- krysa rodu rattus MeSH
- lidé MeSH
- myši MeSH
- ženské pohlaví MeSH
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
The fundamental biochemical processes of 5-methylcytosine (5-mC) synthesis, maintenance, conversion and removal determine the time and spatial pattern of DNA methylation. This has a strong effect on a plethora of physiological aspects of cellular metabolism. While the presence of 5-mC within the promoter region can silence gene expression, its derivative - 5-hydroxymethylcytosine exerts an opposite effect. Dysregulations in the metabolism of 5-mC lead to an altered DNA methylation pattern which is linked with a disrupted epigenome, and are considered to play a significant part in the etiology of several human diseases. A summary of recent knowledge about the molecular processes participating in DNA methylation pattern shaping is provided here.
- MeSH
- 5-methylcytosin metabolismus MeSH
- DNA metabolismus MeSH
- lidé MeSH
- metylace DNA * MeSH
- Check Tag
- lidé MeSH
- Publikační typ
- časopisecké články MeSH
- přehledy MeSH
- Publikační typ
- abstrakt z konference MeSH
PURPOSE: Fruit and vegetable intake is inversely correlated with cancer; thus, it is proposed that an extract of phytochemicals as present in whole fruits, vegetables, or grains may have anti-carcinogenic properties. Thus, the anti-tumour effects of fruit peel polyphenols (Flavin7) in the chemoprevention of N-methyl-N-nitrosourea-induced mammary carcinogenesis in female rats were evaluated. METHODS: Lyophilized substance of Flavin7 (F7) was administered at two concentrations of 0.3 and 3 % through diet. The experiment was terminated 14 weeks after carcinogen administration, and mammary tumours were removed and prepared for histopathological and immunohistochemical analysis. In addition, using an in vitro cytotoxicity assay, apoptosis and proliferation after F7 treatment in human breast adenocarcinoma (MCF-7) cells were performed. RESULTS: High-dose F7 suppressed tumour frequency by 58 % (P < 0.001), tumour incidence by 24 % (P < 0.05), and lengthened latency by 8 days (P > 0.05) in comparison with the control rats, whereas lower dose of F7 was less effective. Histopathological analysis of tumours showed significant decrease in the ratio of high-/low-grade carcinomas after high-dose F7 treatment. Immunohistochemical analysis of rat carcinoma cells in vivo found a significant increase in caspase-3 expression and significant decrease in Bcl-2, Ki67, and VEGFR-2 expression in the high-dose group. Both doses demonstrated significant positive effects on plasma lipid metabolism in rats. F7 significantly decreased survival of MCF-7 cells in vitro in MTT assay by dose- and time-dependent manner compared to control. F7 prevented cell cycle progression by significant enrichment in G1 cell populations. Incubation with F7 showed significant increase in the percentage of annexin V-/PI-positive MCF-7 cells and DNA fragmentation. CONCLUSIONS: Our results reveal a substantial tumour-suppressive effect of F7 in the breast cancer model. We propose that the effects of phytochemicals present in this fruit extract are responsible for observed potent anti-cancer activities.
- MeSH
- antigen Ki-67 genetika metabolismus MeSH
- antitumorózní látky fytogenní analýza farmakologie MeSH
- apoptóza účinky léků MeSH
- experimentální nádory mléčných žláz farmakoterapie MeSH
- flavonoidy analýza farmakologie MeSH
- fragmentace DNA účinky léků MeSH
- kaspasa 3 genetika metabolismus MeSH
- krysa rodu rattus MeSH
- lidé MeSH
- methylnitrosomočovina toxicita MeSH
- MFC-7 buňky MeSH
- modely nemocí na zvířatech MeSH
- ovoce chemie MeSH
- polyfenoly analýza farmakologie MeSH
- proliferace buněk účinky léků MeSH
- protein X asociovaný s bcl-2 genetika metabolismus MeSH
- receptor 2 pro vaskulární endoteliální růstový faktor genetika metabolismus MeSH
- stilbeny analýza farmakologie MeSH
- tyrosin analogy a deriváty metabolismus MeSH
- vztah mezi dávkou a účinkem léčiva MeSH
- zvířata MeSH
- Check Tag
- krysa rodu rattus MeSH
- lidé MeSH
- ženské pohlaví MeSH
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
The effect of dietary administered young barley containing a mixture of phytochemicals to female rats for the prevention of N-methyl-N-nitrosourea-induced mammary carcinogenesis was evaluated. After carcinogen administration (14 wk), mammary tumors were removed and prepared for histopathological and immunohistochemical analysis. Moreover, in vitro evaluation of possible mechanisms in MCF-7 breast cancer cell line was performed. Barley (0.3%) demonstrated mild antitumor effect in mammary carcinogenesis, yet 3% barley did not further improve this effect. Immunohistochemical analysis of rat tumor cells in treated groups showed significant increase in caspase-3 expression and significant reduction in Ki67 expression. In addition, 3% barley significantly decreased dityrosine levels versus control. Barley in higher dose significantly decreased serum low-density lipoprotein-cholesterol in rats. In vitro studies showed that barley significantly decreased survival of MCF-7 cells in 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide assay and significantly decreased 5-bromo-20-deoxyuridine incorporation versus control. Barley prevented cell cycle progression and extended incubation with barley showed significant increase in the percentage of annexin V/propidium iodide-positive MCF-7 cells. Our results propose an antitumor effect for the mixture of phytochemicals present in young barley in a breast cancer model.
- MeSH
- antikarcinogenní látky farmakologie MeSH
- apoptóza MeSH
- experimentální nádory mléčných žláz chemicky indukované patologie prevence a kontrola MeSH
- flavonoidy analýza MeSH
- ječmen (rod) * chemie MeSH
- lidé MeSH
- metabolismus lipidů MeSH
- methylnitrosomočovina toxicita MeSH
- MFC-7 buňky MeSH
- nádory prsu MeSH
- potkani Sprague-Dawley MeSH
- proliferace buněk MeSH
- zvířata MeSH
- Check Tag
- lidé MeSH
- ženské pohlaví MeSH
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
Several neurodegenerative conditions, such as Alzheimer's disease and Parkinson's disease, or vascular dementia and cognitive impairment, are associated with mild hyperhomocysteinemia. Hyperhomocysteinemia is defined as an increase of the homocysteine (Hcy) level beyond 10 microM. Although the adverse effect of Hcy on neurons is well documented, knowledge about the impact of this amino acid on glial cells is missing. Therefore, with the aim to evaluate the neurotoxic properties of Hcy on glial cells, we used a glioblastoma cell line as a study model. The viability of cells was assayed biochemically and cytologically. At a concentration around 50 microM in the culture medium D,L-Hcy induced cell death. It is noteworthy that Hcy induces cell death of human glial cells at concentrations encountered during mild hyperhomocysteinemia. Therefore, we propose that Hcy-induced impairment of neuronal functions along with damage of glial cells may contribute to the etiopathogenesis of neurodegenerative diseases associated with hyperhomocysteinemia.
- MeSH
- buněčná smrt účinky léků fyziologie MeSH
- homocystein farmakologie MeSH
- kultivované buňky MeSH
- lidé MeSH
- nádorové buněčné linie MeSH
- neuroglie účinky léků fyziologie MeSH
- neurony účinky léků fyziologie MeSH
- viabilita buněk účinky léků fyziologie MeSH
- vztah mezi dávkou a účinkem léčiva MeSH
- Check Tag
- lidé MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
Preparáty na báze fosfolipidov sa v klinickej praxi využívajú v rámci prevencie, ale aj ako podporná liečba pri terapii už niekoľko desaťročí. Viaceré štúdie poukazujú na skutočnosť, že biologicky aktívne fosfolipidy (BAF) môžu inhibovať rast niektorých druhov nádorov a ich schopnosť metastázovať. Multiformný glioblastóm (GBM) je u dospelých najčastejším a najagresívnejším druhom mozgového nádoru s prognózou prežívania u novodiagnostikovaných prípadov len niekoľko mesiacov. Štandardná liečba GBM zahŕňa resekciu nádoru, radiačnú terapiu a chemoterapiu temozolomidom (TMZ). V našej práci sme sa zamerali na štúdium vplyvu BAF, obohatenom o 1-O-oktadecyl-2-oleoyl-sn-glycero-3-fosfo-(N-palmitoyl)etanolamín (PNAE), na prežívanie ľudských glioblastómových buniek a jeho schopnosť ovplyvňovať cytotoxický účinok TMZ. Ako experimentálny model pre in vitro štúdium buniek GBM sme použili ľudskú glioblastómovú líniu T98G. Bunky boli inkubované v médiách BAF, bez PNAE (nBAF) a s prítomnosťou PNAE (aBAF) v objemových koncentráciách 0,05–0,3 %. S cieľom overiť vplyv BAF na cytotoxický efekt TMZ, boli bunky predinkubované s aBAF (0,05 %) po dobu 24 hodín a následne inkubované v prítomnosti TMZ. Na kvantifikáciu prežívajúcich buniek sme použili dva typy metód: i) metyl-tiazol tetrazoliový test (MTT) a ii) cytologické farbenie. Naše výsledky ukazujú, že BAF s obsahom PNAE pôsobí cytotoxicky na glioblastómové bunky už pri objemovej koncentrácii 0,05 %. Predinkubácia buniek s aBAF signifikantne potenciuje cytotoxický účinok TMZ na nádorové bunky. Na základe našich výsledkov môžeme tvrdiť, že biologicky aktívne fosfolipidy obohatené o PNAE by mohli byť používané ako podporná zložka pri chemoterapeutickej liečbe GBM s TMZ, zároveň by tak prispeli k zlepšeniu prognózy a kvality života pacientov.
Extracts of biologically active phospholipids (BAP) may exert a positive effect on inhibition of tumor growth and may be used as additive compounds to supplement palliative tumor treatment. The exact mechanisms by which BAP affect the cancer cells are still unknown. Among brain tumors, glioblastoma multiforme (GBM) is considered the most aggressive and the most common primary brain tumor in adults. The treatment of GBM includes surgery, radiotherapy and chemotherapy with an alkylating agent temozolomide (TMZ). With the aim to study the survival of human glioblastoma cells, in media supplemented with BAP alone or in combination with TMZ, we used the cell line T98G as a study model. The effect of BAP on glioblastoma cell survival was assessed by an methyl-thiazol tetrazolium (MTT) assay and also microscopically. Cells were incubated in medium enriched with BAP of different final 1-O-octadecyl-2-oleoyl-sn-glycero-3-phospho-( N-palmitoyl)ethanolamine (PNAE) concentrations in range 0,05 – 0,3 % (aBAP). Our results show that aBAP negatively affected the cell survival, depending on their concentration in culture medium. Pre-culturing of the cells with aBAP for 24 hours before the incubation with TMZ had potentiated the cytotoxic effect of TMZ on GBM cells. The results indicate that BAP with PNAE are capable to either inhibit growth or induce the death of human glioblastoma cells in vitro. In conclusion, the capability of aBAP to potentiate the cytotoxic effect of temozolomide indicates that further testing of aBAP as an adjunct agent to treat GBM, may be of great clinical significance.
- MeSH
- antitumorózní látky alkylující * terapeutické užití MeSH
- chemorezistence MeSH
- dakarbazin antagonisté a inhibitory terapeutické užití MeSH
- fosfolipidethery * farmakokinetika MeSH
- fosfolipidy * farmakokinetika MeSH
- glioblastom * farmakoterapie MeSH
- inhibitory angiogeneze MeSH
- lidé MeSH
- Check Tag
- lidé MeSH
- Publikační typ
- práce podpořená grantem MeSH
