摘要
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。
Objective To assess the effects of p27mt gene transfection on the proliferation and apoptosis 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 ^3Hthymidine incorporation; the cell apoptosis was determined by flow cytometry, TUNEL method and DNA fragmentation analysis. Results The virus titer was 7.95×10^12 cfu/ml, the transduction efficiency was 100 % when multiplicity of infection I〉50, FCM analysis revealed a sub-G, 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.
出处
《肿瘤研究与临床》
CAS
2006年第8期538-540,共3页
Cancer Research and Clinic
基金
湖北省自然科学基金计划资助项目(2005ABA083)
