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腺病毒E1A基因诱导细胞凋亡.E1B19K基因及E1B55K基因抑制细胞凋亡,前者被克隆到腺病毒转移载体pCA13的HCMVIE启动子下游.构建成转移载体pCAE1A。采用lipofectin法将PCAE1A和含腺病毒基因组(E1、E3区缺失)的质粒pBHG11共转染293细胞,7~10d后得到重组病毒v5Ad4。用v5Ad4感染人肺腺癌细胞系A549,结果表明v5Ad4有明显杀伤和裂解肿瘤细胞功能。在人胚肺正常二倍体细胞中,v5Ad4没有表现出可见的细胞毒效应。
Adenovirus E1A gene induces apoptosis. The E1B19K gene and E1B55K gene inhibited apoptosis, and the former was cloned downstream of the HCMVIE promoter of the adenoviral transfer vector pCA13. Constructed as transfer vector pCAE1A. Plasmid pBHG11 containing PCAE1A and adenoviral genome (E1 and E3 regions deleted) was co-transfected into 293 cells by lipofectin method, and recombinant virus v5Ad4 was obtained 7-10 days later. The human lung adenocarcinoma cell line A549 was infected with v5Ad4. The results showed that v5Ad4 significantly kills and lyses tumor cells. In human embryonic lung normal diploid cells, v5Ad4 did not show a visible cytotoxic effect.