癌变·畸变·突变Issue(3)：209-212,4.DOI:10.3969/j.issn.1004-616x.2014.03.011
慢病毒介异的孕烷X受体sh-RNA稳定干扰HepG2细胞株的构建
Establishment of a HepG2 cell line with stable PXR silencing by lentivir us-mediated RNA interference
摘要
Abstract
OBJECTIVE:To establish a HepG2 cell line with stable PXR silencing effect by a lentiviral vector carrying a short hairpin RNA (sh-RNA). METHODS:Three double-stranded sh-RNA targeting the PXR gene were designed,synthesized and cloned into the psi-sH1 sh-RNA vector. The effective recombinant sh-RNA expression plasmid against PXR,which was selected of HEK293T cells with three double-stranded sh-RNA plasmids,was packaged into HEK293T cells and used to infect HepG2 cells after 200 mmol/L puromycin screening for 2-3 weeks. The expression level of PXR was detected by western blot,the CYP3A4 mRNA levels both in sh-RNA PXR HepG2 stable cells and scramble control HepG2 cells were detected by real-time PCR. RESULTS:A recombinant sh-RNA expression plasmid against PXR gene with effective interference was successfully selected and a HepG2 cell line stably transfected with the sh-RNA PXR was established. CONCLUSION:A HepG2 cell line stably transfected with the sh-RNA PXR was established.关键词
人孕烷X受体/原发性肝细胞癌/慢病毒/RNA干扰Key words
human pregnane X receptor/hepatocellular carcinoma/lentivirus/RNA interference分类
医药卫生引用本文复制引用
牛永东,陈理明,吴曼鹏,程訸,徐涵,高分飞,石刚刚..慢病毒介异的孕烷X受体sh-RNA稳定干扰HepG2细胞株的构建[J].癌变·畸变·突变,2014,(3):209-212,4.基金项目
中国博士后科学基金(2012M510200)，广东省自然科学基金(S2012010010955)，广东省医学科研基金(A2013656)，中央财政支持地方高校发展专项资金 （2012M510200）
