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Archive > Volume 17 Issue 3 > 2010,17(3):302-307. DOI:10.3872/j.issn.1007-385X.2010.3.011 Prev Next

Basic Research

Construction of survivinpPRIMEIGF1RmiR30 lentiviral vector and its inhibitory effect on proliferation of liver cancer cells

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    Abstract:
    Objective:To study the effects of survivinpromoterregulated survivinIGF1RmiR30 (surIGFIRmiR30) lentiviral vector on the IGF1R expression and proliferation of hepatoma Hep3B cells. Methods：Survivin promoter was amplified by PCR and surpPRIME plasmid was constructed. Interference sequence targeting IGF1R gene was synthesized and cloned into surpPRIME plasmid, named surIGF1RmiR30. SurIGF1RmiR30, psPAX2, and pMD2G were cotransfected into 293T cells to amplify lentivirus, and the lentivirus titer was examined. IGF1R expression in Hep3B cells was detected by RTPCR and Western blotting analysis, and the proliferation of Hep3B cells was evaluated by CCK8 assay. Results：SurIGF1RmiR30 lentiviral vector regulated by survivin promoter were successfully constructed, and the virus titer was 4.58×109 PFU/ml. Fluorescent protein after surIGF1RmiR30 infection was expressed in Hep3B cells, but not in L02 cells. SurIGF1RmiR30 infection inhibited IGF1R mRNA and protein expressions in Hep3B cells and the proliferation of Hep3B cells, with the inhibitory rate being 60％ at 7 d (P<0.05). Conclusion: SurIGF1RmiR30 lentiviral vector can inhibit IGF1R expression and hepatoma cell proliferation.

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    Project Supported:
    Project supported by the Natural Science Foundation of Jiangsu Higher Institutions (No.09kjb320018)

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