5-氮杂-2’-脱氧胞苷调控RASSF1A基因表达对子宫内膜癌HEC-1-B细胞增殖及凋亡的影响

Effects of 5-Aza-2’-deoxycytidine on proliferation and apoptosis by modulating RASSF1A gene expression in human endometrial carcinoma cell line HEC-1-B

摘要:

目的：探讨甲基化转移酶抑制剂5-氮杂-2’-脱氧胞苷（5-Aza-2’-deoxycytidine，5-Aza-CdR）对人子宫内膜癌（Hu－man endometrial cancer，HEC）-1-B细胞增殖和凋亡的作用及其可能机制。方法：应用不同浓度的 5-Aza-CdR处理HEC-1-B细胞，甲基化特异 PCR（Methylation-specific polymerase chain reaction，MSP）法检测处理前后RAS相关域家族1A（RAS association domain family 1A，RASSF1A）基因甲基化状态，应用 RT-PCR方法检测 RASSF1A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：HEC-1-B细胞 RASSF1A基因启动子区呈高甲基化状态，经过 5-Aza-CdR处理后，RASSF1A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RASSF1A基因甲基化状态，调控RASSF1A基因表达，并能有效地抑制HEC-1-B细胞的增殖和诱导细胞调亡。

Abstract:

Objective：To investigate the effects of 5-aza-2’-deoxycytidine（5-Aza-CdR） on the proliferation and apoptosis in human endometrial cancer（HEC）HEC-1-B cells and the possible mechanism. Methods：HEC-1-B cells were treated with different concentrations of DNA methytransferase inhibitor 5-Aza-CdR. Methylation-specific polymerase chain reaction（MSP） was used to detect pro－moter methylation status of RAS association domain family 1A（RASSF1A） gene and RT-PCR was used to detect re-expression of mRNA. Furthermore，cell growth was estimated by MTT assay and cell apoptosis rate was estimated by flow cytometry. Results：Promoter hypermethylation of the RASSF1A gene was detected in HEC-1-B cells. After treatment with 5-Aza-CdR，the promoter region of the RASSF1A gene exhibited a demethylation status，and RASSF1A gene was re-expressed. Meanwhile，the cellular growth activity decreased and cell apoptosis increased. Conclusions：The 5-Aza-CdR could reverse methylation of RASSF1A gene to regulate the expression of RASSF1A gene，and could effectively inhibit the cellular proliferation and induce apoptosis of endometrial carcinoma cells.