中文摘要：

目的探讨应用整合素连接激酶（integrin-linkedkinase，ILK）特异性siRNA沉默ILK基因的表达对人膀胱癌细胞凋亡的影响。方法将构建好的针对ILK基因的1个特异性siRNA表达质粒和1个无同源性的阴性对照质粒，在脂质体介导下稳定转染人膀胱癌BIU-87细胞，并将实验分为BIU-87siILK、BIU-87vector及BIU-87细胞3组，运用TUNEL凋亡检测试剂盒和流式细胞术检测BIU-87细胞的凋亡，进一步运用Westernblot检测siRNAILK对凋亡相关蛋白Caspase-3、Bax和Bcl-2的影响。结果TUNEL检测结果显示，BIU-87siILK组出现大量凋亡细胞，而BIU-87细胞组和BIU-87vector组仅有少量凋亡细胞出现。流式细胞仪检测结果显示，BIU-87siILK组细胞大约有（75．70±2．00）％的凋亡细胞，而BIU-87vector组和BIU-87细胞组仅有（0．88±0．10）％和（1．66±0．90）％的凋亡细胞，Westernblot结果显示，在BIU．87siILK组中，Bcl-2的表达明显降低（P〈0．05），而Bax和Caspase-3的表达明显升高（P〈0．01）。结论siRNAILK可能通过调节凋亡相关蛋白Bcl-2、Bax及Caspase-3的表达诱导膀胱癌BIU-87细胞凋亡的发生。

英文摘要：

Objective To determine the of human bladder cancer BIU-87 cells. Methods effects of integrin-linked kinase （ILK） siRNA on the apoptosis Two siRNA vectors specific to ILK gene and one non-homol- ogous negative control vector were designed and constructed, then stably transfected into bladder cancer BIU-87 cells via Lipofectamine 2000, and were named as BIU-87 si ILK and BIU-87 vector. Cell apoptosis was assessed by TUNEL kit and flow cytometry. The expression of Caspase-3, Bax and Bcl-2 were assessed by Western blotting. Results The number of TUNEL-positive cells was significantly increased in the BIU-87 si ILK cells when compared with BIU-87 cells and BIU-87 vector cells. Flow cytometry showed that （75.70± 2. 00）% of counted cells became apoptotic in BIU-87 si ILK cells, however, about only （0.88± 0.10）% and （ 1.66 ± 0.90） % of counted cell became apoptotic in BIU-87 vector cells and BIU-87 cells groups respectively. The expression of Bcl-2 was significantly reduced in BIU-87 si ILK cells compared with BIU-87 vector and BIU-87 cells（P 〈 0.05 ）, and that of Caspase-3 and Bax protein was stably increased （P 〈 0.01 ）. Conclusion The siRNA ILK induces apoptosis in bladder cancer BIU-87 cells by regulating apoptosis-related proteins Caspase-3, Bax and Bcl-2.