摘要
目的观察靶向survivin的不同siRNA对EJ28细胞增殖和凋亡的影响。方法通过化学合成方法合成3对siRNA和1对荧光标记的阴性对照siRNA。转染荧光标记的siRNA后,在荧光显微镜下观察转染效率。实验分为siRNA164、siRNA167、siRNA389、阴性对照组、脂质体对照组和细胞对照组。转染后24、48h和72h,MTT法检测siRNA对EJ28细胞增殖的影响,流式细胞术检测凋亡率。结果靶向survivin的序列特异性siRNA均能抑制EJ28细胞的增殖,其中以siRNA164抑制细胞的增殖能力最强(P<0.05),转染48h后的抑制率最高[(41.32±2.54)%];转染48h后,siRNA164组凋亡率最高[(13.20±0.25)%]。结论靶向survivin的RNAi技术在膀胱癌的基因治疗中可能具有一定的价值。
Objective To explore the proliferative and apoptotic effect of 3 different siRNAs targeting survivin in human bladder cancer cell line EJ28. Methods Three siRNAs targeting survivin and 1 fluorescencelabeled siRNA as a negative control were chemically synthesized. Transfetion efficiency was observed under fluorescence microscope after transfecting the fluorescence-labeled siRNAs. EJ28 cells were divided into 6 groups： siRNA164 group, siRNA167 group, siRNA389 group, and negative control group, lipofectin group and cell control group. The proliferation of EJ28 cell was detected by MTT assay at 24, 48 and 72 h after the transfection. Apoptotic rate was detected by Annexin V-FITC-FCM assay at 48 h. Results All the sequence-specific siRNAs targeting survivin efficiently suppressed the proliferation of EJ28 cells, siRNA 164 had the most powerful effect because of its highest inhibitory rate of （41.32 ±2.54 ） % and the highest apoptotic rate of （ 13.20± 0.25 ）% at 48 h after transfection. Conclusion RNAi targeting survivin has a potential value in the gene therapy of bladder cancer.
出处
《第三军医大学学报》
CAS
CSCD
北大核心
2007年第22期2152-2155,共4页
Journal of Third Military Medical University
基金
重庆市卫生局项目(04-2-130)~~
