5-脱氧杂氮胞苷调控p16基因表达对人宫颈癌HeLa细胞增殖及调亡的影响

目的:探讨甲基化转移酶抑制剂5-脱氧杂氮胞苷(5-Aza-CdR)对人宫颈癌HeLa细胞增殖和调亡的作用及其可能机制。方法:应用不同浓度的5-Aza-CdR处理HeLa细胞,甲基化特异PCR(MSP)法检测处理前后p16基因甲基化状态,应用RT-PCR方法检测p16基因mRNA表达,MTT法检测细胞增殖,流式细胞术检测细胞凋亡率。结果:人宫颈癌HeLa细胞p16基因启动子区呈高甲基化状态,经过5-Aza-CdR处理后,p16基因启动子区呈去甲基化状态,其mRNA重新表达,细胞生长受到抑制,细胞凋亡增加。结论:5-Aza-CdR能够逆转p16基因甲基化状态,调控p16基因表达,并有效抑制人宫颈癌HeLa细胞的增殖和诱导细胞调亡。

Effects of 5-Aza-CdR on proliferation and apoptosis by modulating p16 gene expression in human cervical carcinoma cell line HeLa

Objective：To investigate the effects of 5-aza-2-deoxycytidine（5-Aza-CdR） on proliferation and apoptosis in human cervical carcinoma cell line HeLa and the possible mechanism.Methods： HeLa cells was treated with different concentration of DNA methytransferase inhibitor 5-Aza-CdR.Methylation-specific polymerase chain reaction（MSP） was used to detect promoter methylation status of p16 gene and RT-PCR was used to detect re-expression of mRNA.Cell growth was estimated by MTT assay and cell apoptosis rate was determined by flow cytometry.Results： Promoter hypermethylation of the p16 gene was detected in HeLa cells.After treatment with 5-Aza-CdR,the promoter region of the p16 gene exhibited a demethylation status,and the p16 gene mRNA was re-expressed.The cellular growth activity decreased and cell apoptosis increased.Conclusion： The 5-Aza-CdR could reverse the methylation of p16 gene to regulate the expression of p16 gene,and could effectively inhibit the cellular proliferation and induce apoptosis of cervical carcinoma cells.