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采用同源重组方法构建RA5 3 8cDNA重组体腺病毒 ,通过转导人卵巢癌细胞系SK OV 3和人黑色素瘤细胞系WM 983A ,证实腺病毒可高效转导RA5 3 8基因 ,能显著抑制肿瘤细胞的生长 ,其抑制率分别达 85 %和 73 % ;并显著降低两细胞系的集落形成能力 .对两细胞系的裸鼠皮下移植瘤进行治疗实验结果表明 ,RA5 3 8能明显抑制肿瘤的生长 .流式细胞计数和DNA片段化分析证实 ,RA5 3 8可引起肿瘤细胞G1期阻滞和 /或凋亡 .对c myc蛋白的Westernblot分析发现 ,RA5 3 8有明显下调c myc基因表达的作用 .根据这些结果 ,我们认为RA5 3 8重组体腺病毒有可能成为有应用价值的肿瘤基因治疗药物 .
RA53-8 cDNA recombinant adenovirus was constructed by homologous recombination method. Through transduction of human ovarian cancer cell line SK OV 3 and human melanoma cell line WM 983A, it was confirmed that adenovirus can efficiently transduce RA5 38 gene and can significantly inhibit tumorigenesis. The growth rate of the cells was up to 85% and 73%, respectively, and the colony-forming ability of the two cell lines was significantly reduced. The results of the treatment experiments on the nude mice subcutaneously transplanted tumors of two cell lines showed that RA53 8 can significantly inhibit tumors. Growth, flow cytometry, and DNA fragmentation analysis confirmed that RA53 8 can cause G1 phase arrest and/or apoptosis in tumor cells. Western blot analysis of c myc protein revealed that RA53 8 significantly down-regulated the expression of c-myc gene. Based on these results, we believe that the RA5-38 recombinant adenovirus may become an oncogenic gene therapy drug with application value.