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目的 研究一种结合肿瘤基因治疗与病毒治疗优势的新型肿瘤治疗载体系统 ,即基因 病毒载体系统。方法 利用病毒重组技术将抗癌基因插入肿瘤细胞特异性的增殖病毒基因组中 ,通过细胞病理作用、荧光显微镜、免疫酶链技术及电镜等技术分别观察病毒的杀伤效应、报告基因绿色荧光蛋白、抗癌基因小鼠白细胞介素 12表达量及病毒复制情况。结果 构建了一种新型基因 病毒载体系统 ,该载体系统腺病毒E1b 5 5kDa蛋白缺失 ,保留了腺病毒E1a蛋白。该载体系统具有肿瘤增殖病毒ONYX 0 15相似功能 ,即它可在肿瘤细胞内复制及增殖 ,而在正常细胞内不能复制及增殖 ,从而特异性杀灭肿瘤细胞。它还具有更大的优势 ,即该载体系统可携带各种抗癌基因以进一步提高抗肿瘤的疗效。应用该载体系统携带该报告基因绿色荧光蛋白可使绿色荧光蛋白在肿瘤细胞内高效表达 ,其表达量明显高于传统基因治疗的腺病毒载体系 ,而在正常细胞内低表达 ,与传统腺病毒载体系统相似或更低。应用该载体系统携带抗癌基因小鼠白细胞介素 12 ,也产生类似结果 ,并在电镜中证实该载体系统携带抗癌基因小鼠白细胞介素 12可在肿瘤细胞株中复制及增殖。结论 基因 病毒载体是将抗癌基因插入肿瘤增殖病毒基因组 ,通过肿瘤增殖病毒在肿瘤细胞内特异性复制?
OBJECTIVE: To study a novel tumor therapy vector system that combines the advantages of tumor gene therapy and virus therapy, ie, gene viral vector system. Methods The anti-cancer gene was inserted into the tumor cell-specific proliferating virus genome by virus recombination technique. The killing effect of the virus was observed by cytopathology, fluorescence microscopy, enzyme-linked immunosorbent assay and electron microscopy. The expression of interleukin - 12 in murine gene and the replication of virus. Results A novel gene viral vector system was constructed, in which the adenovirus E1b 5 5 kDa protein was deleted and the adenovirus E1a protein was retained. The vector system has the similar function as the tumor proliferating virus ONYX 0 15, that is, it can replicate and proliferate in tumor cells, but can not replicate and proliferate in normal cells to specifically kill tumor cells. It has the added advantage that the vector system can carry a variety of anti-oncogenes to further enhance antitumor efficacy. Using this vector system to carry the reporter green fluorescent protein, EGFP can be efficiently expressed in tumor cells, and its expression level is obviously higher than that of the traditional gene therapy adenovirus vector system, but it is low in normal cells. Compared with the traditional adenovirus Carrier systems are similar or lower. Using this vector system to carry the anti-oncogene mouse interleukin 12 also produced similar results, and confirmed by electron microscopy that the vector system carrying the anti-oncogene mouse interleukin 12 can replicate and proliferate in tumor cell lines. Conclusions Gene viral vectors insert the anti-oncogene into the tumor-proliferating virus genome and replicate specifically within the tumor cells by the tumor-proliferating virus.