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siRNA沉默整合素连接激酶对人膀胱癌细胞凋亡的影响 被引量：2

Integrin-linked kinase siRNA induces apoptosis in human bladder cancer BIU-87 cells
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摘要 目的探讨应用整合素连接激酶(integrin-linked kinase,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-87 si ILK、BIU-87 vector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Western blot检测siRNA ILK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-87 si ILK组出现大量凋亡细胞,而BIU-87细胞组和BIU-87 vector组仅有少量凋亡细胞出现。流式细胞仪检测结果显示,BIU-87 si ILK组细胞大约有(75.70±2.00)%的凋亡细胞,而BIU-87 vector组和BIU-87细胞组仅有(0.88±0.10)%和(1.66±0.90)%的凋亡细胞,Western blot结果显示,在BIU-87 si ILK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 siRNA ILK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。 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.
作者 高娟 康炜 陈俊霞 朱军 潘湘阳
机构地区 西安医学院附属医院检验科 重庆医科大学基础医学院细胞生物学与遗传学教研室
出处 《第三军医大学学报》 CAS CSCD 北大核心 2012年第21期2154-2157,共4页 Journal of Third Military Medical University
基金 国家自然科学基金面上项目(81071719) 重庆市自然科学基金(CSTC2009BB5273)~~
关键词 整合素连接激酶 膀胱癌 凋亡 integrin linked kinase bladder cancer apoptosis
分类号 R394.2 [医药卫生—医学遗传学] R730.23 [医药卫生—肿瘤]
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参考文献15

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二级参考文献47

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第三军医大学学报
2012年 第21期

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