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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。
OBJECTIVE: To construct the adenovirus CNHK200-mIFN-γ and proliferation-deficient adenovirus AdEasy-mIFN-γ, and to compare the expression of mIFN-γ protein with CNHK200-mIFN-γ, ONYX-015 and wild Adenovirus Ad5 in normal and tumor cells in the proliferation of further observation of its anti-tumor effect. Methods: The proliferative ability of the virus in the cells was detected by virus proliferation assay. The proliferation and replication of CNHK200-mIFN-γ in Hep3B cells were observed by transmission electron microscopy. The expression of mIFN-γ was detected by the virus-infected tumor cells. The killing effect. Results: CNHK200-mIFN-γ and ONYX-015 only proliferated in tumor cells, and the former had stronger proliferative ability. The expression of mIFN-γ in CNHK200-mIFN-γ cells was closely related to the proliferation of tumor cells, while the expression of mIFN-γ in AdEasy-mIFN-γ cells was almost undetectable. ONYX-015 and CNHK200-mIEN-γ are non-lethal to normal cells, but effectively kill cancer cells. Conclusion: The insertion of exogenous gene mIFN-γ did not change the selective proliferation of proliferating virus in tumor cells. Proliferating adenovirus CNHK200-mIFN-γ has good tumor selectivity and proliferation, and can effectively express mIFN-γ, and effectively kill cancer cells, showing its good clinical application prospects.