慢病毒siRNA靶向干扰YAP基因胃癌细胞株的建立  

Establishment of Gastric Cancer Cells with Stable Interference by siRNA Targeting YAP Gene

作　　者：周洲[1] 朱金水[1] 许志朋[1] 

机构地区：[1]上海交通大学附属第六人民医院消化科,上海200233

出　　处：《现代生物医学进展》2011年第4期605-610,共6页Progress in Modern Biomedicine

基　　金：上海市科委动物实验基金重点项目(10140902500)

摘　　要：目的:构建并鉴定YAP基因短发夹干扰RNA(shRNA)慢病毒载体,建立稳定干扰YAP基因表达的胃癌细胞株SGC7901。方法:荧光定量PCR检测YAP基因在多种胃癌细胞株中的表达情况。构建重组靶向YAP基因的shRNA慢病毒表达质粒PGC-shRNA-YAP,用脂质体转染的方法将载体导入胃癌细胞。经杀稻瘟菌素筛选后,建立稳定表达siRNA的细胞株。荧光定量PCR检测干扰效率。结果:在胃癌细胞株SGC7901中,YAP基因显示高表达。测序验证PGC-shRNA-YAP重组质粒构建成功。将重组质粒稳定转染入胃癌细胞株SGC7901后能明显抑制YAPmRNA表达水平。结论:成功构建了PGC-shRNA-YAP慢病毒重组质粒,建立了靶向稳定干扰YAP基因表达的siRNA胃癌细胞株SGC7901。Objective：To construct siRNA expression vector targeting YAP gene and establish the gastric cancer cells SGC7901 with stable inhibition of YAP gene.Methods：Rea-time PCR was used to examine the YAP gene expression in sevel gastric cancer cell lines.The recombinant lentivirus PGC-shRNA-YAP vector was constructed,which was transfected into gastric cancer cells by Lipofec-tamine 2000.After gastric cancer cells with constant expression of the PGC-shRNA-YAP were established by blasticidin selection,the interfere efficicncy of YAP mRNA expression level was detected by real-time PCR.Results：There was high expression of YAP gene in the gastric cancer cell line SGC-7901.Recombinant lentivirus PGC-shRNA-YAP vector was successfully constructed by the identifica-tion of sequencing.The recombinant vector markedly inhibited the expression of YAP gene in gastric cancer cell line SGC-7901.Conclusion：Recombinant lentivirus vector PGC-shRNA-YAP was successfully constructed and gastric cancer cell line SGC7901 with stable siRNA expression targeting YAP gene was established.

关 键 词：YAP基因 慢病毒载体 胃癌细胞株SGC7901 RNA干扰 

分 类 号：Q75[生物学—分子生物学] Q78