SARDH/PIPOX converts exogenously added sarcosine while providing methyl groups to methyl donor S-adenosyl methionine

Kód výsledku v IS VaVaI

<a href="https://www.isvavai.cz/riv?ss=detail&h=RIV%2F62156489%3A43210%2F17%3A43911932" target="_blank" >RIV/62156489:43210/17:43911932 - isvavai.cz</a>

Nalezeny alternativní kódy

RIV/00216305:26620/17:PU125541

Výsledek na webu

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DOI - Digital Object Identifier

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Jazyk výsledku

angličtina

Název v původním jazyce

SARDH/PIPOX converts exogenously added sarcosine while providing methyl groups to methyl donor S-adenosyl methionine

Popis výsledku v původním jazyce

Prostate cancer (PCa) is the most common cancer among men worldwide1. Sarcosine, an N-methyl derivative of the amino acid glycine, plays a substantial role in PCa aggressiveness and progression as its levels correlate tightly with its invasiveness2. In our experiment, three PCa cell lines (non-malignant PNT1A, metastatic LNCaP and malignant 22Rv1) were incubated with sarcosine (10 µM) for 24 h and the results of indirect immunofluorescence showed significant (p &lt; 0.05) increase in expression, particularly of enzymes involved in the conversion of sarcosine to glycine - sarcosine dehydrogenase (SARDH) and l-pipecolic acid oxidase (PIPOX), compared to only negligible effect on expression of glycine N-methyltransferase (GNMT) and dimethylglycine dehydrogenase (DMGDH). Such conversion provides free methyl group for methyl donor S-adenosyl methionine (SAH) to be converted to S-adenosyl homocysteine (SAH). Changes in DNA methylation are pivotal aspect during cancer initiation and progression and are present in variety of cancers including PCa1,3. Analyses of global DNA methylation of sarcosine-supplemented cells revealed time-depended increase, with the highest levels found after 72 h incubation. Bisulphite-sequencing polymerase chain reaction was further used to validate the promoter methylation of 13 candidate genes, which are responsible for gene expresion regulation, cell cycle control, DNA repair and signal transduction (namely TP53, JUN, FOS, AR, CCND2, CDKN2A, GPX3, KRAS, MYCN, GSTP1, RBP1, EDNRB and CD44). Finally, using wound-healing assay, we show that using demethylation agent Azacitidine, sarcosine-induced invassiveness can be inhibited, which highlights the pivotal role of sarcosine in methylation status of prostate cells.

Název v anglickém jazyce

SARDH/PIPOX converts exogenously added sarcosine while providing methyl groups to methyl donor S-adenosyl methionine

Popis výsledku anglicky

Prostate cancer (PCa) is the most common cancer among men worldwide1. Sarcosine, an N-methyl derivative of the amino acid glycine, plays a substantial role in PCa aggressiveness and progression as its levels correlate tightly with its invasiveness2. In our experiment, three PCa cell lines (non-malignant PNT1A, metastatic LNCaP and malignant 22Rv1) were incubated with sarcosine (10 µM) for 24 h and the results of indirect immunofluorescence showed significant (p &lt; 0.05) increase in expression, particularly of enzymes involved in the conversion of sarcosine to glycine - sarcosine dehydrogenase (SARDH) and l-pipecolic acid oxidase (PIPOX), compared to only negligible effect on expression of glycine N-methyltransferase (GNMT) and dimethylglycine dehydrogenase (DMGDH). Such conversion provides free methyl group for methyl donor S-adenosyl methionine (SAH) to be converted to S-adenosyl homocysteine (SAH). Changes in DNA methylation are pivotal aspect during cancer initiation and progression and are present in variety of cancers including PCa1,3. Analyses of global DNA methylation of sarcosine-supplemented cells revealed time-depended increase, with the highest levels found after 72 h incubation. Bisulphite-sequencing polymerase chain reaction was further used to validate the promoter methylation of 13 candidate genes, which are responsible for gene expresion regulation, cell cycle control, DNA repair and signal transduction (namely TP53, JUN, FOS, AR, CCND2, CDKN2A, GPX3, KRAS, MYCN, GSTP1, RBP1, EDNRB and CD44). Finally, using wound-healing assay, we show that using demethylation agent Azacitidine, sarcosine-induced invassiveness can be inhibited, which highlights the pivotal role of sarcosine in methylation status of prostate cells.

Druh

O - Ostatní výsledky

CEP obor

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OECD FORD obor

10608 - Biochemistry and molecular biology

Projekt

<a href="/cs/project/GA16-18917S" target="_blank" >GA16-18917S: Studium metabolismu sarkosinu a jeho participace na vývoji nádorů prostaty</a><br>

Návaznosti

P - Projekt vyzkumu a vyvoje financovany z verejnych zdroju (s odkazem do CEP)

Rok uplatnění

2017

Kód důvěrnosti údajů

S - Úplné a pravdivé údaje o projektu nepodléhají ochraně podle zvláštních právních předpisů