siRNA介导的survivin基因沉默诱导人膀胱癌BIU87细胞凋亡的研究.docVIP

siRNA介导的survivin基因沉默诱导人膀胱癌BIU-87细胞凋亡的研究 作者:曹正国,诸禹平,周四维 作者单位：1.安徽省立医院，安徽 合肥 230001；2. 华中科技大学同济医学院附属同济医院，湖北 武汉 430030 【摘要】 　　［目的］ 研究siRNA（small interference RNA）对人膀胱癌BIU-87细胞凋亡和survivin基因表达的影响。［方法］ 利用Ambion公司设计软件，设计并体外转录合成针对survivin基因的3个特异性siRNA，通过脂质体将siRNA转入BIU-87细胞，分别采用MTT和DNA原位末端标记（TUNEL）法检测siRNA对BIU-87细胞生长抑制率（IR）和凋亡指数（AI），半定量逆转录聚合酶链反应(RT-PCR)和Western Blotting检测siRNA对survivin mRNA及其蛋白表达的影响。［结果］转染后的siRNA1～3组的BIU-87细胞的IR（41.84%、56.21%、54.13%）和AI（21.98%、36.54%、31.34%）均分别显著高于正常对照组（1.98%和3.17%）（P＜0.05），survivin mRNA及其蛋白表达水平均显著低于对照组；其中siRNA2～3对BIU-87细胞的IR、AI和survivin表达的抑制作用均显著高于siRNA1。［结论］体外转录合成的siRNA可抑制BIU-87细胞survivin的表达，诱导BIU-87细胞凋亡，从而抑制BIU-87细胞生长，为siRNA介导的膀胱肿瘤基因沉默提供实验依据。 【关键词】 siRNA　RNAi　膀胱肿瘤　survivin　细胞凋亡　基因沉默 　　A Study SiRNA Induced Survivin Gene Silencing on Cell Apoptosis in Human Bladder Cancer Cell Line BIU-87 in vitro CAO Zheng-guo1, ZHU Yu-ping1, ZHOU Si-wei2, et al. (1.Anhui Provincial Hospital, Hefei 230001, China; 2.Tongji Hospital, Tongji Medical College, Huazhong University of Science and Technology, Wuhan 430030, China) Abstract: ［Purpose］ To investigate the effects of siRNA (small interference RNA) on the cell apoptosis and survivin gene expression in human bladder cancer cell line BIU-87 in vitro. ［Methods］ With the software of Ambion Corporation, 3 specific siRNA targeted survivin gene were synthesized after designed and transcription in vitro. After siRNA were transferred into BIU-87 cells by liposome, MTT and TUNEL methods were used to detect the proliferation inhibitory rate (IR) and apoptosis index (AI) of BIU-87 cells, semi-quantitive reverse transcription polymerase chain reaction (RT-PCR). Western Blotting techniques were used to examine the effects of siRNA on the expressions of the survivin gene and protein. ［Results］ The IR （41.84%, 56.21% and 54.13%）and AI (21.98%, 36.54% and 31.34%) of BIU-87 cell in the siRNA 1～3 group were respectively significantly higher than those in the co