GPC3-siRNA真核质粒载体的构建及Huh-7细胞转染  

Construction of GPC3-siRNA eukaryotic plasmid vector and its transfection of Huh-7 cells

作　　者：温继育[1] 邹文俊[2,3] 刘忠考[2] 黎然[2] 邱志东[4] 李明意[4] 缪辉来[4] 

机构地区：[1]广东医学院附属医院肿瘤科,广东湛江524001 [2]广东医学院附属医院肝胆外科研究室,广东湛江524001 [3]解放军第422医院普外二区,广东湛江524009 [4]广东医学院附属医院肝胆外科,广东湛江524001

出　　处：《广东医学院学报》2014年第2期143-146,共4页Journal of Guangdong Medical College

基　　金：广东省科技计划项目(No.20101700501)

摘　　要：目的探讨GPC3-siRNA真核质粒载体构建并转染Huh-7细胞的可行性。方法构建真核质粒载体pcDNA3.1+GPC3和GPC3-siRNA-1633,采用脂质体法瞬时转染至Huh-7细胞中。MTT法检测转染后Huh-7细胞的细胞活性,荧光定量PCR、Western blot分别检测GPC3 mRNA和蛋白表达。结果 GPC3-siRNA-1633基因和GPC3真核质粒载体转染Huh-7细胞后细胞存活率高,转染GPC3-siRNA-1633的Huh-7细胞出现GPC3 mRNA和蛋白表达下调。结论 GPC3-siRNA-1633可成功转染至Huh-7细胞,并抑制GPC3表达。Objective To investigate the construction of GPC3-siRNA eukaryotic plasmid vector and its transfection of Huh-7 cells. Methods The eukaryotic plasmid vectors pcDNA3.1＋GPC3 and GPC3-siRNA-1633 were constructed, and then transfected into Huh-7 cells using liposome transient transfection method. Cell viability and GPC3 mRNA and protein expression in Huh-7 cells were determined by MTT method, fluorescence quantitative PCR and Western blot, respectively. Results The survival rate of Huh-7 cells was high after transfection with the eukaryotic plasmid vectors pcDNA3.1＋GPC3 and GPC3-siRNA-1633. GPC3 mRNA and protein expression of Huh-7 cells was down-regulated after transfection with GPC3-siRNA-1633 eukaryotic plasmid vector. Conclusion GPC3-siRNA-1633 eukaryotic plasmid vector can be transfected into Huh-7 cells, and inhibit the GPC3 expression.

关 键 词：GPC3 真核质粒 转染 HUH-7细胞 

分 类 号：R735.7[医药卫生—肿瘤]