【摘要】 目的:构建增殖型腺病毒CNHK200-mIFN-γ和增殖缺陷型腺病毒AdEasy—mIFN-γ,比较两者在肿瘤细胞中表达mIFN-γ蛋白的能力以及CNHK200-mIFN-γ,ONYX-015及野生型腺病毒Ad5在正常及肿瘤细胞中的增殖力,进一步观察其抗瘤效果。方法:以病毒增殖试验检测病毒在细胞中的增殖能力;透射电镜观察CNHK200-mIFN-γ在Hep3B细胞中的增殖复制;检测病毒感染肿瘤细胞后mIFN-γ的表达;CPE实验观察增殖病毒对细胞的杀伤效应。结果:CNHK200-mIFN-γ和ONYX-015仅在肿瘤细胞中增殖,且前者增殖力较强。CNHK200-mIFN-γ在肿瘤细胞中表达mIFN-γ,且表达量与病毒增殖密切相关,而AdEasy—mIFN-γ在肿瘤细胞中的mIFN-γ表达几乎不能测到。ONYX-015与CNHK200-mIEN-γ对正常的细胞无杀伤性,但能有效地杀伤癌细胞。结论:外源基因mIFN-γ的插入没有改变增殖病毒在肿瘤细胞中选择性增殖的特性。增殖型腺病毒CNHK200-mIFN-γ具有良好的肿瘤选择性及增殖性,且可以有效表达mIFN-γ,并且有效杀伤癌细胞,显示其良好的临床应用前景。更多还原

【Abstract】 Objectives: To construct the replication-competent adenovirus CNHK200-mIFN-γ and the replication-deficient adenovirus AdEasy-mIFN-γ, and to compare their expression of mIFN-γ and the replicative activities of CNHK200-mIFN-γ,ONYX-015 and wide-type adenovirus ( Ad5) in normal and cancer cells. Their antitumor activities were also observed. Methods: The replicative activities of viruses in cells were measured by viral replication assay; The replication of CNHK200-mIFN-γ in Hep3B cells was observed under the electron microscopy; The expression of mIFN-γ in cancer cells was detected by ELISA. CPE assay was used to detect viral antitumor activity. Results: Both CNHK200-mIFN-7 and ONYX-015 replicated only in cancer cells, and CNHK200-mIFN-γ replicated more potential than ONYX-015; The infection of CNHK200-mIFN--y led to an obvious expression of mIFN-γ, whereas the expression of mIFN-γ mediated by AdEasy-mlFN-γ could not be detected at all. CNHK200-mIFN-γ and ONYX-015 could kill cancer cells but spare normal cells. Conclusions: The insertion of mIFN-γ causes no alterations on the tumor-specific replication of the original virus. CNHK200-mIFN-γ can selectively replicate and effectively express mIFN-γ in tumor cells, and specifically kill cancer cells, suggesting a splendid future as a new anticancer agent.更多还原