携带hIFN-γ的增殖型腺病毒治疗胃癌的体外实验研究

In vitro Potent Antitumor Activity of the Oncolytic Adenovirus Expressing hIFN-γ in Gastric Cancer

目的:研究增殖型腺病毒CNHK300-hIFN-γ在胃癌细胞中的增殖、杀伤能力与表达hIFN-γ蛋白的能力。方法:病毒增殖实验比较CNHK300-hIFN-γ与CNHK300在胃癌细胞SGC-7901与成纤维细胞BJ中的增殖能力;ELISA法比较CNHK300-hIFN-γ和复制缺陷型腺病毒Ad-hIFN-γ在细胞中hIFN-γ蛋白的表达量;细胞活力MTT检测法与CPE实验观察CNHK300-hIFN-γ对胃癌细胞与正常细胞的杀伤效应。结果:增殖实验结果表明CNHK300-hIFN-γ能选择性地在端粒酶阳性的胃癌细胞中大量增殖;ELISA实验结果表明CNHK300-hIFN-γ在胃癌细胞中hIFN-γ的表达量可至117.26±15.92ng/ml;MTT与CPE实验表明CNHK300-hIFN-γ对胃癌细胞有显著的杀伤性,对正常细胞无明显杀伤。结论:增殖型腺病毒CNHK300-hIFN-γ感染胃癌细胞后增殖活跃并能大量表达目的基因hIFN-γ,对胃癌细胞有明显杀伤作用但对正常细胞无明显杀伤,为胃癌的病毒基因治疗进一步提供了理论依据。

Objective： To compare the expression of hIFN-γ, replicative activities and anfitumor activities of CNHK300 - hIFN-γ and Ad - hIFN-γ in normal and gastric cancer cells. Method： The replicative activities of viruses in cells were measured by viral replication assay; CPE assay was used to detect the antitumor effect of viruses; The expression level of hIFN-γ in cancer cells was detected by ELISA. Result： The infection of CNHK300- hIFN-γ led to an obvious expression of hIFN-γ in gastric cancer ceils. The vector system CNHK300- hIFN-γpossessed more replicated potential than Ad - hIFN -γ and could specifically kill most of SGC - 7901 cells at MOI value of 0.1, which was much better than that by the traditional adenoviral vector. Conclusion： CNHK300- hIFN-γ can selectively replicate and effectively express hIFN-γ in tumor cells, and specifically kill gastric cancer cells, suggesting a splendid future as a new anticancer agent.