5-杂氮脱氧胞苷逆转膀胱癌细胞WIF-1基因转录表达的研究

目的 探讨去甲基化药物5-杂氮脱氧胞苷对膀胱癌WIF-1基因表达的影响及其生物学效应.方法 用5-杂氮脱氧胞苷处理膀胱癌细胞株T24和BIU87,RT-PCR检测WIF-1 mRNA表达情况,免疫细胞化学和Western blot检测WIF-1蛋白表达变化,甲基化特异性PCR检测WIF-1启动子甲基化状态变化,MTT法检测5-杂氮脱氧胞苷对膀胱癌细胞的增殖抑制作用,TUNEL法和流式细胞仪检测细胞凋亡率.结果 5-杂氮脱氧胞苷处理72 h后,WIF-1 mRNA及蛋白表达明显增加,而且恢复为启动子未甲基化状态,膀胱癌细胞增殖明显抑制,FCM法检测T24和BIU87的凋亡率分别为(18.2±2.6)%和(17.3±2.7)%.结论 DNA甲基化是导致WIF-1不表达的重要原因,5-杂氮脱氧胞苷可以通过逆转WIF-1甲基化状态而恢复WIF-1表达,从而抑制膀胱癌细胞的生长并诱导细胞凋亡.5-杂氮脱氧胞苷有望成为膀胱癌的有效辅助化疗药.

Apoptosis and growth inhibition of bladder cancer cells induced by 5-aza-2'-deoxycyfidine via reactivating Wnt inhibitor factor-1 gene expression

Objective To study the effect of 5-aza-2'-deoxyeytidine on WIF-1 gene expression in bladder cancer and its inhibitory effect on bladder cancer cells.Methods The change of WIF-1 mRNA and protein expression,methylation status of WIF-1 gene promoter,inhibitory rate and apoptotie rate were detected bv RT-PCR,immunoeytoehemistry,Western blot,methylation-specific PCR,MTT and flow cytometry,respectively.Results After 5-aza-2'-deoxyeytidine treatment on bladder csncer lines,T24 and BIU87 for 72 h,WIF-1 mRNA and protein expression was increased significantly,and WIF-1 gene promoter become unmethylated.The growth of bladder cancer cells was inhibited significantly and the apoptotic rates was(18.2±2.6)%and(17.3±2.7)%for T24 and BIU87,respectively.Conclusion DNA methylation Was an important factor of silencing expression of WIF-1.could inhibit growth and induce apoptosis of bladder cancer cells through reversing unmethvlation status of bladder cancer.5-aza-2'-deoxycytidine would be an effective aajunctive ehemotherapeutic agent for bladder cancer.