重组腺病毒介导的p^16与p^53基因联合转移对人肺癌细胞系 …

目的 探讨ｐ＾１６和ｐ＾５３基因对肺癌细胞的协同抑制效应及凋亡诱导作用。方法 首先用同源重组技术构建重组ｐ＾１６和ｐ＾５３腺病毒载体,然后单独或联合感染人肺癌细胞系Ｈ３５８,用免疫组织法及Ｗｅｓｔｅｒｎ ｂｌｏｔ检测腺病毒介导的基因转移效率与表达水平,用克隆形成实验、原位末端标记及流式术观察它们对Ｈ３５８生长特性及凋亡的影响,结果 免疫组织化学染色结果表明重组腺病毒载体可高效地将外源基因ｐ＾５３转

Effects of adenovirus-mediated p16 and p53 genes transfer on apoptosis and cell cycle of lung carcinoma cells

Objective To explore the synergistic inhibition effect and apoptosis induction of p16 and p53 genes on lung carcinoma cells. Methods E1 deficient and replication defective recombinant p16 and p53 adenoviruses were generated by liposome mediated co transfection of recombinant plasmid pAdCMV p16 or pAdCMV p53 along with pJM17 and homologous recombination in 293 packaging cell. The lung cancer cell line H358, which had a homozygous deletion of p53 gene and no expression of p16 mRNA and protein, was infected with recombinant p16 and p53 adenovirus either individually or together. Results Immunohistochemical analysis showed that recombinant adenovirus could transfer p53 gene into tumor cell with 98% efficiency. Western blot indicated that p16 and p53 proteins were expressed at a high level in infected H358 cell. Inhibition effect of p53 gene on proliferation of H358 cell was weaker than that of p16 gene, and the combined use of both genes could completely prevent the proliferation of H358 cell. In situ end labeling and flow cytometry indicated that p16 could result in G 1 arrest of cell cycle and did not induce H358 cells to undergo apoptosis; p53 also induced apoptosis of few cells besides G 1 arrest; and the simultaneous use of p16 and p53 genes could induce marked apoptosis of H358 cells. Conclusion p16 and p53 genes possess the synergistic inhibiting effect on growth of lung cancer cells and can cooperate to induce apoptosis of H358 cell. The combined application of recombinant p16 and p53 adenoviruses can be used as a new strategy for cancer gene therapy.