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

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Prostate cancer (PCa) is the most common cancer among men worldwide (1). 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 invasiveness(2). 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 < 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 PCa (1,3). 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.

Prostate cancer (PCa); sarcosine

MICHÁLEK, P.; STRMISKA, V.; LACKOVÁ, Z.; KŘÍŽKOVÁ, S.; BUCHTELOVÁ, H.; GURÁŇ, R.; POMPEIANO VANÍČKOVÁ, L.; DOSTÁLOVÁ, S.; ZÍTKA, O.; ADAM, V.; HEGER, Z.

7. 10. 2017

1107-3756

INTERNATIONAL JOURNAL OF MOLECULAR MEDICINE

40

1

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