5-氮杂脱氧胞苷联合孕激素对子宫内膜癌细胞株HEC-1-B生长的影响

Study on the effect of 5-Aza-CdR and progesterone on PR-B negative endometrial cancer cell lines HEC-1-B

目的:研究5-氮杂脱氧胞苷(5-Aza-CdR,ADC)联合孕激素对孕激素受体B亚型(PR-B)缺失的子宫内膜癌细胞株HEC-1-B增殖、凋亡、细胞周期分布的影响及其机制。方法:5-Aza-CdR、醋酸甲羟孕酮(medroxyprogesterone acetate,MPA)单独及联合作用于HEC-1-B细胞后,运用MTT比色法及流式细胞仪分别检测各组细胞的生长、凋亡及细胞周期分布情况;采用RT-PCR技术、甲基化特异性PCR(MSP)分别检测HEC-1-B细胞中PR-B mRNA表达及PR-B基因启动子的甲基化状态。结果:(1)5-Aza-CdR对HEC-1-B细胞的生长抑制作用呈剂量和时间依赖性,联合用药组的抑制作用更为明显(P<0.01),且使细胞阻滞在G1期;(2)5-Aza-CdR以剂量依赖的方式上调PR-B mRNA的表达(P<0.01);(3)5-Aza-CdR可以抑制PR-B基因启动子的甲基化状态。结论:(1)5-Aza-CdR联合孕激素能明显抑制HEC-1-B细胞增殖,促进其凋亡,二者联用可加强MPA对细胞周期的阻滞作用;(2)5-Aza-CdR能逆转PR-B基因甲基化而使其恢复表达,从而增强孕激素的作用。

Objectives：To study the effect of 5-Aza-CdR combined with progesterone on proliferation of Endometrial cancer cell line HEC-1-B cells and hypermethylation of PR-B gene.Methods：The effect of 5-Aza-CdR and/or MPA on proliferation,apoptosis and cell cycle of HEC-1-B cell was separately studied by MTT and flow cytometry.The effect of 5-Aza-CdR on the expression and methylation status of PR-B gene of HEC-1-B cell was separately observed by RT-PCR and MSP.Results：（1） The growth inhibition of 5-Aza-CdR on HEC-1-B cell line was concentration-dependent and time-dependent and the united group presented a more prominent proapoptotic effects（P0.01）,inhibited the cells at G1 period.（2） 5-Aza-CdR can increase the expression of PR-B mRNA by a concentration-dependent manner（P0.01）.（3） The hypermethylation of PR-B promoter can be inhibited by 5-Aza-CdR.Conclusions：（1） 5-Aza-CdR combine with progesterone can significant inhibit HEC-1-B cell growth and promote cells apoptosis,and the combination of this two medicine can enhance the effect of MPA on the cell-cycle,block the cells at G1 period.（2） 5-Aza-CdR can renew the expression of PR-B gene,so it can enhance the effect of progesterone.