重组腺病毒介导突变型p27基因转移对肝癌细胞SMMC-7721生长的影响

 目的 探讨突变型p27基因（p27mt）对肝癌细胞SMMC-7721的细胞增生和凋亡的调节作用。方法 利用重组体腺病毒Ad-p27mt转染培养的人肝癌细胞SMMC-7721，用3H -TdR掺入法检测细胞增生；流式细胞术、DNA片段分析法、TUNEL法检测细胞凋亡。结果 重组体腺病毒Ad-p27mt在MOI≥ 50时， 可达到100 %的转导效率。Ad-p27mt转染肝癌细胞后，3H -TdR掺入检测发现细胞增生抑制；流式细胞术检测在G1期前出现亚二倍体凋亡峰；细胞DNA抽提电泳后发现凋亡特征性梯带。Ad-p27mt组及空白对照组TUNEL法检测凋亡指数分别为58.6±4.3及4.5±1.6，差异有统计学意义（P＜0.01）。结论 重组腺病毒介导的p27mt基因转移可抑制肝癌SMMC-7721细胞增生并诱导其凋亡。

Effects of p27mt gene transfection mediated by replication deficient adenovirus on the proliferation and apoptosis of human hepatocellular carcinoma cells

Objective To assess the effects of p27mt gene transfection on the proliferation and apop－tosis of human hepatocellular carcinoma cell (HCC) lines SMMC-7721. Methods A replication deficient adenovirus vector encoding p27mt （Ad-p27mt） was used and p27mt cDNA was transfected into human SMMC-7721 cell lines in vitro. The synthesis of DNA in SMMC-7721 cells was determined by using 3H-thymidine incorporation; the cell apoptosis was determined by flow cytometry, TUNEL method and DNA frag－mentation analysis. Results The virus titer was 7.95×1012 cfu/ml, the transduction efficiency was 100 % when multiplicity of infection ≥50, FCM analysis revealed a sub-G1 cell peak in Ad-p27mt transduced hepa－tocellular carcinoma cell lines. Agarose electrophoresis showed marked ladder .The difference of apoptotic in－dex between the Ad-p27mt group and the control group was statistically significant (58.6±4.3, vs 4.5±1.6, P <0.01). Conclusions p27mt gene could inhibit the proliferation and increase the apoptosis of human HCC cells.