目的：探讨survivin启动子调控的重组慢病毒survivinpPRIMEIGF1RmiR30载体(简写为surIGF1RmiR30)对肝癌Hep3B细胞IGF1R表达和细胞生长的影响。方法：PCR扩增survivin启动子，构建surpPRIME；将针对〖STBX〗IGF1R〖STBZ〗基因的干扰序列与surpPRIME载体连接，构建surIGF1RmiR30慢病毒载体。将surIGF1RmiR30、psPAX2和pMD2G质粒共转染293T细胞，扩增慢病毒，检测病毒滴度。以surIGF1RmiR30感染人肝癌Hep3B细胞和胎肝L02细胞，RTPCR、Western blotting检测Hep3B细胞IGF1R的表达，CCK8法检测Hep3B细胞的生长。结果：成功构建survivin启动子调控的慢病毒载体surIGF1RmiR30，滴度为4.58×109 PFU/ml。SurIGF1R miR30感染后，Hep3B细胞中特异表达荧光蛋白，L02细胞中基本没有表达。SurIGF1RmiR30感染Hep3B细胞可阻断IGF1R mRNA和IGF1R蛋白的表达。SurIGF1RmiR30感染抑制肝癌细胞的生长，第7天时的抑制率达60％(P<0.05)。结论：成功构建的重组慢病毒载体surIGF1RmiR30 可有效地降低肝癌细胞中IGF1R的表达，抑制肝癌细胞的增殖。

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.

江苏省高校自然科学研究基金项目（No.09kjb320018）