siRNA沉默Wip1基因对人肝癌HepG2细胞的影响  

Effect of the down-regulation Wip1 gene on HepG2 cells

作　　者：赵殿堂 李光兵[1] 李哲[1] 张允成[1] 刘军[1] 

机构地区：[1]山东大学附属省立医院器官移植肝胆外二科,山东济南250021

出　　处：《中国现代普通外科进展》2015年第1期1-5,共5页Chinese Journal of Current Advances in General Surgery

基　　金：国家自然科学基金面上项目(81373172)

摘　　要：目的 :研究Wip1对肝癌细胞生物学活性的影响。方法:设计并合成能沉默Wip1基因的siRNA质粒,通过脂质体介导转染肝癌HepG2细胞,并分为实验组和空白对照组。用Western blot和qRT-PCR检测转染后细胞内Wip1基因的表达水平,CCK-8检测HepG2细胞增殖,用划痕实验和Transwell实验检测细胞迁移及侵袭能力的改变。结果:将沉默Wip1基因的质粒导入肝癌HepG2细胞后,与空白对照组相比,Wip1的蛋白和mRNA表达水平明显降低(P<0.05);CCK-8检测结果显示转染后第4天细胞增殖能力较空白对照组降低(P<0.05),转染后第5天细胞增殖能力较空白对照组明显降低(P<0.01);实验组迁移及侵袭细胞数均少于空白对照组(P<0.05)。结论:靶向设计的siRNA能有效降低Wip1蛋白和mRNA的表达水平,抑制HepG2细胞的增殖并降低细胞的迁移及侵袭力。Objective： To study the effects of Wip1 on the biological activity of hepatocellular carcinoma cells. Methods： Design and synthesize specific siRNA, which can inhibit the expression of Wip1 gene. Transfect HepG2 cells with siRNA using lipofectamine 2000 and experiment were divided into experimental group and control group. The protein and m RNA expression level of Wip1gene were studied using Western blot and q RT-PCR. The proliferation of HepG2 cells was detected using CCK-8 and cell migration and invasion capability were tested by scratch test and Transwell assay. Results： The expression of Wip1 protein and m RNA were significantly decreased in siRNA group compared with the control group（P〈0.05）. CCK-8 testing results showed that cell proliferation was reduced compared with untransfected group after four days transfection（P〈0.05）and significantly reduced after five days transfection（P〈0.01）. Transwell results show that cell number were significantly less than that of non transfected group（P〈0.05）. Conclusion：Specific siRNA was successfully designed and can effectively reduce the protein and m RNA expression level of Wip1, which can inhibit the growth of HepG2 cells and reduced cell migration and invasion.

关 键 词：肝肿瘤 Wip1基因 SIRNA 细胞生物学特性 

分 类 号：R735.7[医药卫生—肿瘤]