Lentivirus vectors construction of SiRNA targeting interference GPC3 gene and its biological effects on liver cancer cell lines Huh-7

查看全文 作　　者：Chang-Jiang [1]Lei;Chun [2]Yao;Qing-Yun [3]Pan;Hao-Cheng [1]Long;Lei [1]Li;Shu-Ping [4]Zheng;Cheng [1]Zeng;Jian-Bin [1]Huang

机构地区：[1]Department of General Surgery,the Second Affiliated Hospital of Jianghan University(Wuhan City Fifth Hospital);[2]Wuhan Institute of Hematology,the Second Affiliated Hospital of Jianghan University(Wuhan City Fifth Hospital);[3]Integrated Department,Branch of Jianqiao.Hanyang Affiliated Hospital of Wuhan University of Science and Technology;[4]Department of Pediatrics,the Second Affiliated Hospital of Jianghan University(Wuhan City Fifth Hospital)

出　　处：《Asian Pacific Journal of Tropical Medicine》索引2014年第7卷第10期,共7页

基　　金：supported by Wuhan Municipal Science and Technology Bureau of applied basic research project(No.2013062301010823);Wuhan City health planning medieal research project(No.WX14A11) 摘　　要：Objective:To build GPC3 gene short hairpin interference RNA(shRNA)slow virus veclor.observe expression of Huh-7 GPC3 gene in human liver cell line proliferation apoptosis and the effect of GPC3 gene influencing on liver cancer cell growth,and provide theoretical basis for genc therapy of liver cancer.Methods:Hepatocellular carcinoma cell line Huh-7 wsa transfected by a RNA interference technique.GPC3 gene expression in a variety of liver cancer cell lines was detected by fluorescence quantitative PCR.Targeted GPC3 gene seqnences of small interfering RNA(siRNA)PGC-shRNA-GPC3 were restructured.Stable expression cell linse of siRNA were screened and established with the heplp of liposomes(lipofectamine^(TM2000))as carrier transfcetion of human liver cell lines.In order to validate siRNA interference efficiency.GPC3 siRNA mRNA expression was detected after transfection by using RT-PCR and Western blot.The absorbance value of the cells of blank group,untransfection group and transfection group,the cell cycle and cell apoptosis were calculated,and effects of GPC3 gene nn Huh-7 cell proliferation and apoptosis were observed.Results:In the liver cancer cell lines Huh-7 GPC3 gene showed high expression.PGC-shRNA-GPC3 recombinant plasmid was constructde successfully via sequencing validation.Stable recombinant plasmid transfected into liver cancer cell linse Huh-7can obviously inhibit GPC3 mRNA expression level.Conclusions:The targeted GPC3 siRNA can effectively inhibit the expression of GPC3. 关键词：GPC3 GENE SLOW virus CARRIER Liver cancer cell LINES RNA INTERFERENCE

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Lentivirus vectors construction of SiRNA targeting interference GPC3 gene and its biological effects on liver cancer cell lines Huh-7

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