Effects of 5-Aza-2′-deoxycytidine on proliferation of human breast cancer cell line Hs578T and methylation of PRDM10 gene / 天津医药

ABSTRACT

Objective To investigate effects of 5-Aza-2′-deoxycytidine (5-Aza-CdR) on proliferation of human breast cancer cell line Hs578T,and the methylation status of PRDM10 gene in vitro in this cell line.Methods The human breast cancer cell line Hs578T was cultured with 1,3 and 5 μmol/L DNA methylation inhibitor 5-Aza-CdR respectively, and untreated cells were used as control.Cell proliferation was detected by MTT assay.Methylation-Specific PCR(MSP)was used to detect the methylation status of PRDM10 gene. The mRNA and protein expression levels of PRDM10 gene were detected by RT-PCR and Western blot assay. Results MTT results showed that the higher the concentration of 5-Aza-CdR,and the longer the treatment time,the more significant inhibitory effect on the proliferation of Hs578T cells.Compared with the control group(0 μmol/L),the proliferation of Hs578T was significantly inhibited after the treatment for 72 h in the 1 μmol/L group, and for 48 h in the 3 μmol/L and 5 μmol/L groups (P<0.05). MSP results showed that the higher the concentration of 5-Aza-CdR,the more significant demethylation of PRDM10.Results of RT-PCR and Western blot showed that the higher the concentration of 5-Aza-CdR, the higher the expression levels of mRNA and protein in PRDM10 (P<0.05).Conclusion 5-Aza-CdR could inhibit the cell proliferation of Hs578T,which might be related to the demethylation of PRDM10 gene in the cells.