慢病毒介异的孕烷X受体sh-RNA稳定干扰HepG2细胞株的构建

doi:10.3969/j.issn.1004-616x.2014.03.011

Abstract

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.

Key words: human pregnane X receptor, hepatocellular carcinoma, lentivirus, RNA interference

NIU Yong-dong，CHEN Li-ming，WU Man-peng，CHENG He，XU Han，GAO Fen-fei，SHI Gang-gang. Establishment of a HepG2 cell line with stable PXR silencing by lentivirus-mediated RNA interference[J]. Carcinogenesis, Teratogenesis & Mutagenesis, doi: 10.3969/j.issn.1004-616x.2014.03.011.

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Establishment of a HepG2 cell line with stable PXR silencing by lentivirus-mediated RNA interference

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