山东医药2016，Vol.56Issue(11)：1-4,4.DOI:10.3969/j.issn.1002-266X.2016.11.001
转染沉默IGF1R基因的肝癌细胞株增殖、迁移及侵袭能力观察
Construction of RNAi lentiviral vector targeting IGF1R gene and its effect on proliferation, migration and invasion of hepatocellular carcinoma cells
摘要
Abstract
Objective To design and screen an effective small interfering RNA (siRNA) targeting human insulin-like growth factor 1 receptor (IGF1R) gene, to construct the siRNA lentiviral vector , and to observe its effect on the prolifera-tion, migration and invasion of hepatocellular carcinoma cells .Methods According to siRNA design principle , three siR-NAs targeting IGF1R gene and one negative control siRNA were designed and synthesized .They were transfected into Huh 7 cells.IGF1R mRNA expression levels in Huh7 cells were detected by real-time fluorescent quantitative PCR at 24 h after transfection, and we screened the most effective siRNA .After lentiviral expression vector was constructed , it was transfect-ed into 293T cells and packed into lentiviral .Huh7 and Hep3B cells were infected with the pLVX-shRNA2-IGF1R_002 lentiviral to screen the stable cell lines .The stable cell lines were cultured in large .We detected the changes in the IGF 1R mRNA expression , cell proliferation , cell migration and invasion .Results The real-time fluorescent quantitative PCR showed that IGF1R_002 showed the highest inhibition effect (relative inhibition rate 78.6%) at the concentration of 100 nmol/L.The expression levels of IGF1R mRNA in Huh7 and Hep3B cells with the lentiviral were significantly reduced , the proliferation abilities were inhibited , and the abilities of cell migration and invasion were significantly decreased as com-pared with those of Huh7 and Hep3B cells without any virus infection and Huh7 and Hep3B cells infected with pLVX-shR-NA2 empty vector (all P＜0.01).Conclusions The most effective siRNA targeting human IGF1R gene, namely IGF1R_002, is screened out.The most effective siRNA-mediated IGF1R gene silencing can significantly suppress the prolifera-tion, migration and invasion of Huh 7 and Hep3B cells.关键词
RNA干扰/人胰岛素样生长因子1受体基因/小干扰RNA/肝细胞癌Key words
RNA interference/human insulin-like growth factor 1 receptor gene/small interfering RNA/hepatocellu-lar carcinoma分类
医药卫生引用本文复制引用
别彩群,黄秋燕,颜英,石珩,汤绍辉..转染沉默IGF1R基因的肝癌细胞株增殖、迁移及侵袭能力观察[J].山东医药,2016,56(11):1-4,4.基金项目
深圳市科技计划研究项目(JCYJ20140414160300592). （JCYJ20140414160300592）
