[Demethylation in the promoter region of MTHFR gene and its mRNA expression in cultured human vascular smooth muscle cells induced by homocysteine].
OBJECTIVE
To investigate the effect of homocysteine (Hey) on the methylation modification of 5, 10- methylenetetrahydrofolate reductase(MTHFR) gene promoter and its mRNA expression in cultured human vascular smooth muscle cells (HVSMCs).


METHODS
HVSMCs from umbulical arteries were isolated and cultured in DMEM/F12 medium, which clinic-relevant and very high homocysteine concentrations (0,30,100,200,500 and 1000 micromol/L) were added for 72h treatment. Methylation-specific polymerase chain reaction(MS-PCR) was performed to catalyze the methylation pattern of the MTHFR promoter region, semiquantitative RT-PCR was used to detect the MTHFR mRNA expression after Hcy treatment.


RESULTS
The homocysteine induced demethylation in the promoter region of MTHFR gene at all concentrations of Hcy, the methylation pattern in MTHFR gene displayed significant hypomethylation and the MTHFR mRNA expression demonstrated obvious upregulation as well.


CONCLUSIONS
The homocysteine could induce demethylation in the promoter region of MTHFR gene in HVSMCs and upregulate the MTHFR mRNA expression.
Wang
Lizhen
L
West China College of Preclinic Medical Sciences and Forensic Medicine, Sichuan University, Chengdu 610041, China. wanglizhen1979@163.com
Zhang
Jingge
J
Wang
Shuren
S
D002478:Cells, Cultured; D006710:Homocysteine / Q000494:pharmacology*; D006801:Humans; D008745:Methylation; D042965:Methylenetetrahydrofolate Reductase (NADPH2) / Q000235:genetics / Q000378:metabolism*; D009131:Muscle, Smooth, Vascular / Q000166:cytology / Q000378:metabolism*; D011401:Promoter Regions, Genetic / Q000235:genetics*; D012333:RNA, Messenger / Q000235:genetics / Q000378:metabolism; D014469:Umbilical Arteries / Q000166:cytology
D012333:RNA, Messenger; D006710:Homocysteine; D042965:Methylenetetrahydrofolate Reductase (NADPH2)