Preliminary study of NDRG1 gene promoter methylation status in prostate cancer / 中华泌尿外科杂志

ABSTRACT

Objective To evaluate the methylation status of prostate cancer NDRG1 gene promoter region,and to explore the influence of methylation inhibitor 5-azacytidine on NDRG1 gene's mRNA expression in prostate cancer cells and its effects on cell proliferation.Methods Bisulfite-sequencing PCR (BSP) were used to detect the NDRG1 gene promoter methylation status in prostate cancer and BPH tissue,prostate cancer cell lines (PC3,22RV1,LNCaP and DU145) and human normal prostate cell line's RWPE-1.After 10 μmol/L 5-azacytidine were used on LNCaP and DU145 cells for 72 h,5-azacytidine's influence on cell proliferation was analyzed with MTT,two prostate cancer cell lines NDRG1 mRNA expressions were detected with RT-PCR.Results The methylation rates of NDRG1 gene in prostate cancer cell lines PC-3,22RV1,LNCaP and DU145 were (24.8 ± 3.3) ％,(36.2 ± 2.5) ％,(48.6 ± 2.8) ％ and (69.5 ± 1.7) ％,respectively.Methylation rate of Human normal prostate cell lines RWPE-1 was (4.8 ± 4.5) ％;prostate carcinoma was (48.6 ± 5.3) ％,BPH tissue was (4.3 ± 2.1) ％.The differences between groups were statistically significant.After 10 μmol/L 5-azacytidine added on LNCaP and DU145 cells for 72 h,NDRG1 gene demethylation occurred in both cells,its mRNA expression enhanced 8-9 times compared with previous and its cell growth was inhibited (P ＜ 0.05).Conclusions NDRG1 gene promoter region's hypermethylation is one of the reasons of its aberrant expression in prostate cancer.5-azacytidine can reverse NDRG1 gene promoter methylation status,regulate the expression of the gene and can inhibit prostate cancer cell proliferation.