双调控双功效溶瘤腺病毒的构建及意义

Construction and identification of dual-regulated oncolytic adenovirus with dual-efficacy

目的构建表达Survivin基因小发卡RNA和内皮抑素基因的双调控双功效肿瘤特异性溶瘤腺病毒(CNHK500-shSRV-mE,以下简称双调控腺病毒),为肝癌的基因治疗奠定基础。方法以人端粒酶逆转录酶启动子(hTERT)调控腺病毒E1a基因,缺氧调控元件序列(HRE)调控E1b基因,重组双调控双功效的肿瘤特异性溶瘤腺病毒载体;于E1区插入Survivin基因序列设计特异shRNA和全长内皮抑素基因构建双调控腺病毒。将双调控腺病毒分别感染人肝癌细胞株SMMC-7721、BEL-7402,观察病毒的增殖活性。结果双调控腺病毒能在肝癌细胞中特异性高拷贝增殖,而在正常细胞系中几乎不增殖。结论双调控腺病毒成功构建;其为肝癌的基因治疗奠定了基础。

Objective To construct the dual-regulated oncolytic adenovirus carrying surviving-shRNA and endostatin in order to investigate its replicative activity in hepatocellular carcinoma. Methods The hTERT promoter was cloned and used to control Ela expression. HRE was cloned and used to control Elb expression. The survivin shRNA and mouse en- dostatin gene were inserted into adenoviral genome. The recombinant adenovirus was used to infect hepatocellular carcinoma cell lines and its replicative activity was observed. Result The recombinant adenovirus CNI-IKS00-shSRV-mE was replica- ted selectively in cancer cells but not normal cells. Conclusion CNHKS00-shSRV-mE can replicate selectively in cancer cells, which can be used in cancer gene therapy.