survivin特异性RNAi表达载体的构建及其干扰效果鉴定  

Construction and effect identification of RNAi eukaryotic expression vectors of survivin*

作　　者：汪欣[1] 杨军[1] 许波[1] 熊宇[1] 张从纪[1] 

机构地区：[1]第三军医大学西南医院口腔科,重庆400038

出　　处：《重庆医学》2009年第14期1760-1761,F0002,共3页Chongqing medicine

基　　金：重庆市自然科学基金资助项目(CSTC;2007BB5064)。

摘　　要：目的构建survivin基因的RNAi真核表达载体并观察其对转染腺样囊性癌细胞株ACC-2后survivin表达变化以及对细胞凋亡的影响。方法设计、合成siRNA(small interfering RNA),构建表达载体pGenesil-shRNA-survivin,将重组质粒导入人腺样囊性癌ACC-2细胞株。采用逆转录-聚合酶链反应(RT-PCR)法检测转染后的ACC-2细胞中survivin mRNA表达的变化,用TUNEL检测细胞凋亡的变化。结果经测序模版序列和设计序列完全正确,在pGenesil-shRNA-survivin转染ACC-2细胞株中,survivin mRNA表达明显降低,转染的ACC-2细胞凋亡显著增加。结论成功构建了survivin基因的RNAi真核表达载体,且其对腺样囊性癌细胞有一定的干扰效果。Objeetive To investigate apoptosis effect of siRNA on survivin in Adenoid cystic carcinoma cells lines. Methods siRNA was designed and synthesized. Eukaryotic expression vector pGenesil-shRNA-survivin was constructed. Recombinant Plasmid was transfected into ACC-2 cells. The expression of survivin mRNA was detected by RT-PCR and cell apoptosis was detected by TUNEL. Results The recombinant plasmid was successfully constructed. The expression of survivin mRNA was obviously reduced. ACC-2 cells apoptosis was promoted. Conclusion The RNAi eukaryotic vector pGenesil-shRNA-survivin was constructed successfully. The RNAi inhibitory effect on theACC-2 cells is confirmed.

关 键 词：SURVIVIN 质粒 RNAI 

分 类 号：R73-36[医药卫生—肿瘤]