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  

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

作　　者：傅斌[1] 郎斌[2] 徐华[2] 马鑫[2] 张军[2] 许凯[2] 艾星[2] 王保军[2] 周辉霞[2] 张国玺[2] 王超[2] 史涛坪[2] 居正华[2] 张旭[2] 

机构地区：[1]南昌大学第一附属医院泌尿外科,330006 [2]华中科技大学同济医学院附属同济医院泌尿外科

出　　处：《中华实验外科杂志》2008年第3期337-339,F0003,共4页Chinese Journal of Experimental Surgery

基　　金：国家自然科学基金资助项目（30571858、30701010）

摘　　要：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124和BIU87，RT-PCR检测WIF-1 mRNA表达情况，免疫细胞化学和Westernblot检测WIF一1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72h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。Objective To study the effect of 5-aza-2′-deoxycytidine 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 apoptotic rate were detected by RT-PCR, immunocytochemistry, Western blot, methylation-specific PCR, MTT and flow cytometry, respectively. Results After 5-aza-2′-deoxycytidine treatment on bladder cancer 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 unmethylation status of bladder cancer. 5-aza-2′- deoxycytidine would be an effective adjunctive chemotherapeutic agent for bladder cancer.

关 键 词：膀胱癌 甲基化 基因表达 

分 类 号：R737.14[医药卫生—肿瘤] R979.1[医药卫生—临床医学]