survivin基因沉默对腺样囊性癌细胞生长和凋亡的影响.docVIP

survivin基因沉默对腺样囊性癌细胞生长和凋亡的影响 汪 欣，杨 军，许 波，熊 宇，张从纪 （第三军医大学西南医院口腔科，重庆 400038） 摘 要： 目的 探讨RNA干扰下调survivin基因表达后腺样囊性癌细胞生物学行为变化及其与caspase-3基因表达的相关性。方法 设计、合成small interfering RNA，siRNA），构建表达载体pGenesil-shRNA-urvivin，将重组质粒导入人腺样囊性癌ACC-细胞株 。采用逆转录-聚合酶链反应(RT-PCR)、Western blot法分别检测转染后的ACC-细胞中urvivin、caspase-3 mRNA及蛋白表达的变化用噻唑蓝(MTT)法检测细胞的增殖(TUNEL)法及透射电子显微镜研究细胞凋亡。结果 测序证实表达载体构建成功，在pGenesil-shRNA-urvivin的ACC-细胞株中 , urvivin mRNA及蛋白表达明显降低 ,mRNA及蛋白表达明显ACC-2细胞增殖受到抑制。结论urvivin基因的RNAi真核表达载体，pGenesil-shRNA-survivin重组质粒能有效抑制腺样囊性癌细胞。RNA干扰; 腺样囊性癌细胞株; urvivin; caspase-3 中图法分类号： 文献标识码：A The Effect of survivin Gene Silencing in Adenoid cystic carcinoma cells lines on Growth and Apoptosis WANG Xin,YANG Jun,XU Bo,XIONG Yu,ZHANG Cong-ji （Department of oral, First Affiliated Hospital,Third Military Medical University, Chongqing 400038, China） Abstract: Objective To investigate the influence to the biological behavior of Adenoid cystic carcinoma cells after downregulation of survivin and the relationship of expression of caspase-3. Methods We designed and synthesized siRNA .Eukaryotic expression vector pGenesil-shRNA-survivin was constructed .Recombinant Plasmid was transfected into ACC-2 cells, The positive clones were selected with G418 .The expression of survivin and caspase-3 mRNA and protein was detected by RT-PCR and Western blot.The propagation of Adenoid cystic carcinoma cells was detected by MTT. Cell apoptosis was detected by TUNEL staining and transmission electronmicroscope. Results The recombinant plasmid was successfully constructed .The expression of survivin mRNA and protein was obviously reduced . The expression of caspase-3 mRNA and protein was obviously increased. ACC-2 cells propagation was inhibited. The cells showed typical apoptotic characteristics. Conclusion The RNAi eukaryotic vector pGenesil-shRNA-survivin was constructed successfully.Recombinant plasmid pGenesil-shRNA-survivin can specially suppress adenoid cystic carcin