摘要
本研究利用逆转录病毒载体LXSN构建了插入突变的膜结合型TNF-α基因的重组逆转录病毒载体，将重组载体及空载体导入包装细胞CRIP．筛选G418抗性克隆。测定病毒滴度均为1×10^4CFU／ml，并转入胃癌细胞MGC-803．经筛选分别获得抗性克隆MGC-803^T及MGC-803^T,经Southern杂交证实目的基因已整合在MGC-803^T细胞基因组中。MGC-803^T对L929细胞有杀伤作用．而对照细胞无杀伤作用。MGC-803^T细胞的形态，体外增殖率无明显改变。裸鼠体内接种结果表明，与对照组相比，MGC-803^T细胞在裸鼠体内的生长受到明显抑制，表现为肿瘤发生延缓．肿瘤重量明显减轻。上述结果表明．逆转录病毒介导的突变型膜结合型TNF-α基因可在人胃癌细胞中获得有效表达．并能改变其生物学特性。
We used retrovial vector LXSN to construct recombinant retroviral vector with mutant membrane bound TNF - α gene. The vectors were introduced into packaging cell line, CRIP cells. The G418-resistant colonies were selected and the supernatants of the colonies were used to determine the virus titers. The titer of virus was 1× 104CFU/ml and the retroviral vectors were used to tranduced the human gastric cancer cell line, MGC-803 cells. The results of southern blot assay showed thai the targel gene had integrated into the genomic DNA of MGC-803T. MGC - 803Tcells were ablc to kill L929 cell line, but the parent cell line showed no cytotoxicily to the cells at all. There was no any variance in the morphological appearance and growlh rate in vilro of MGC-803N and MGC-803T cells. The results of inoculation in nude mice with these cells indicated that MGC-803T cells showed a considerable decrease in size of tumor. These results suggested that the retroviral vectors expressing mulant TNF -α gene were successfully construed. MGC - 803T cells showed cytotoxicily slrongly lo L929 cell line in vilro and lumorigenicity .
出处
《中国肿瘤生物治疗杂志》
CAS
CSCD
1996年第1期17-20,共4页
Chinese Journal of Cancer Biotherapy
