5-氮杂-2′-脱氧胞苷对人乳腺癌细胞Hs578T增殖和PRDM10基因甲基化的影响

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

目的分析5-氮杂-2′-脱氧胞苷(5-Aza-Cd R)对体外培养的人乳腺癌细胞Hs578T增殖的影响,及其对此细胞中PRDM10基因甲基化状态的影响。方法分别采用浓度为0、1、3、5μmol/L的5-Aza-Cd R处理体外培养的人乳腺癌细胞系Hs578T。MTT实验检测细胞增殖情况,甲基化特异性PCR(MSP)法检测PRDM10的甲基化状态;RT-PCR和Western blot分别检测PRDM10的m RNA和蛋白表达水平。结果 MTT结果显示,5-Aza-Cd R的浓度越高、处理时间越长,对Hs578T细胞增殖的抑制作用越显著。与0μmol/L组比较,1μmol/L组处理72 h后及3、5μmol/L组处理48 h后,Hs578T的增殖受到显著抑制(P<0.05)。MSP结果显示,5-Aza-Cd R的浓度越高,对PRDM10的去甲基化作用越显著。RT-PCR和Western blot结果显示,5-Aza-Cd R的浓度越高,PRDM10的m RNA和蛋白表达水平越高(P<0.05)。结论 5-Aza-Cd R可抑制Hs578T细胞的增殖,可能与该细胞中PRDM10基因的去甲基化作用有关。

To investigate effects of5-Aza-2′-deoxycytidine(5-Aza-CdR)on proliferation of human breast cancer cell line Hs578T was cultured with1,3and5μmol/L DNA methylation inhibitor5-Aza-CdR respectively,and untreated cells were used as control.Cell proliferation was detected by MTT assay.Methylation-Specific PCR(MSP)wasused to detect the methylation status of PRDM10gene.The mRNA and protein expression levels of PRDM10gene weredetected by RT-PCR and Western blot assay.Results MTT results showed that the higher the concentration of5-Aza-CdR,and the longer the treatment time,the more significant inhibitory effect on the proliferation of Hs578T cells.Comparedwith the control group(0μmol/L),the proliferation of Hs578T was significantly inhibited after the treatment for72h in the1μmol/L group,and for48h in the3μmol/L and5μmol/L groups(P<0.05).MSP results showed that the higher theconcentration of5-Aza-CdR,the more significant demethylation of PRDM10.Results of RT-PCR and Western blot showedthat the higher the concentration of5-Aza-CdR,the higher the expression levels of mRNA and protein in PRDM10(P<0.05).Conclusion5-Aza-CdR could inhibit the cell proliferation of Hs578T,which might be related to the demethylationof PRDM10gene in the cells.