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目的：研究逆转录病毒介导干扰素基因转移及在肝癌细胞中特异表达作用。方法：将人β干扰素（ＨｕＩＦＮ－β）ｃＤＮＡ克隆到逆转录病毒载体ＰＬ位点，分别构建了转录受ＳＶ４０启动子驱动的载体ＭＮＳＩＦＮＢ和转录受人甲胎蛋白增强子（ＡＦＰｅ）调控的载体ＭＮＡＩＦＮＢ。用脂质体转染法将载体分别转导ＰＡ３１７包装细胞，质粒转染率为每１０５ＰＡ３１７细胞（４～２５）×１０３ＣＦＵ／μｇ，病毒感染率（４．５～５００）×１０４ＣＦＵ／ｍｌ。重组病毒在４μｇ／ｍｌｐｏｌｙｂｒｅｎｅ存在条件下感染人肝癌、肾癌及黑色素瘤细胞。结果：ＮｅｏＲ克隆ＲＮＡ斑点杂交及ＩＦＮ活性分析证明，人ＡＦＰｅ可促进异源启动子启动ＨｕＩＦＮ－β基因在合成ＡＦＰ的人肝癌细胞中高效特异转录和表达。结论：ＡＦＰ＂持家基因＂顺式调控序列在合成ＡＦＰ的肝癌细胞中特异驱动ＩＦＮ－β基因表达，该研究为肝癌特异性免疫基因治疗提供了资料。
Objective: To study retrovirus-mediated interferon gene transfer and specific expression in hepatoma cells. METHODS: The human interferon-β (HuIFN-β) cDNA was cloned into the retroviral vector PL site, and the vector MNSIFNB driven by the SV40 promoter transcription and the vector regulated by the human alphafetoprotein enhancer (AFPe) transcription were constructed. MNAIFNB. The PA317 packaging cells were transduced with liposome transfection method. The transfection rate of the plasmid was 105 317 CFU/μg per 105 PA317 cells and the virus infection rate was 4.5 to 500×104 CFU/ml. The recombinant virus infected human liver cancer, kidney cancer and melanoma cells in the presence of 4 μg/ml polybrene. RESULTS: NeoR clone dot blot hybridization and IFN activity analysis demonstrated that human AFPe could promote heterologous promoter-initiated HuIFN-beta gene transcription and expression in human hepatoma cells synthesized with AFP. Conclusion: The cis-regulatory sequence of AFP “housekeeping gene” specifically drives the expression of IFN-β gene in AFP-producing hepatocellular carcinoma cells. This study provides data for hepatoma-specific immune gene therapy.