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为探讨肿瘤坏死因子基因对肝癌的治疗作用,分别构建SV40早期启动子和白蛋白增强子/启动子调控小鼠肿瘤坏死因子基因的逆转录病毒载体MNST和MNAT,以及SV40早期启动子和人甲胎蛋白增强子/SV40启动子调控的逆转录病毒载体LTSN和LTASN。构建物导入PA317细胞中包装成重组病毒,感染肿瘤细胞,证明白蛋白增强子/启动子和甲胎蛋白增强子均能使肿瘤坏死因子基因在肝癌细胞中高效特异表达。MNAT病毒和PA317/MNAT产病毒细胞invivo法基因治疗有特异抗肝癌效果。提示组织特征蛋白“持家基因”转录调控序列在肿瘤组织特异性免疫基因治疗研究中有重要意义。
To investigate the therapeutic effect of tumor necrosis factor gene on hepatocellular carcinoma, the retroviral vectors MNST and MNAT, which regulate the mouse tumor necrosis factor gene by the SV40 early promoter and albumin enhancer/promoter, and the SV40 early promoter and human A, respectively, were constructed. The fetal protein enhancer/SV40 promoter regulates the retroviral vectors LTSN and LTASN. The construct was introduced into PA317 cells and packaged into a recombinant virus to infect tumor cells, demonstrating that both the albumin enhancer/promoter and the alpha fetoprotein enhancer enable efficient and specific expression of tumor necrosis factor genes in hepatoma cells. MNAT virus and PA317/MNAT producing virus cells in vivo gene therapy have specific anti-liver cancer effect. These results suggest that the “regulatory gene” transcriptional regulatory sequences of tissue characteristic proteins have important significance in the study of tumor-specific immune gene therapy.