逆转录病毒介导PTEN基因抑制人肝癌细胞系的生长

Inhibitory effects of retrovirus mediated PTEN gene on human hepatocellular carcinoma cell line

目的 研究外源性 PTEN基因对人肝癌细胞系HHCC的细胞周期、增殖及裸鼠致瘤能力的影响 ,探索肝癌基因治疗新途径 .方法 将携有 PTEN基因的真核表达载体体外转染 HHCC细胞 ,筛选稳定转染的细胞克隆并扩增培养 ,以免疫组化方法检测 PTEN基因的表达情况 ,应用流式细胞仪、电子显微镜、生长曲线测定等方法研究 PTEN基因对肝癌细胞周期、超微结构、生长速率等特性的影响 ,并观察导入人 PTEN基因对 HHCC细胞裸鼠致瘤能力的影响 .结果 稳定转染 PTEN基因的 HHCC肝癌细胞中 PTEN蛋白稳定表达 ,细胞周期明显改变 ,细胞从 G1 期到 S期发生抑制 ,细胞超微结构有退行性改变 ,细胞增殖活性下降 70 .6 %裸鼠致瘤能力抑制率 72 .2 % .结论 转染外源性 PTEN基因可阻止HHCC肝癌细胞由 G1 期进入 S期 ,并抑制肿瘤细胞增殖 .

AIM To investigate the suppressive effects of exogenous PTEN gene on HHCC hepatocellular carcinoma cell line and to probe new methods of gene therapy for hepatocellular carcinoma. METHODS A eukaryotic expression vector containing PTEN gene was transfected into human HHCC cell under mediation of lipofectamine, and positive cell clones were selected and amplified. Expression of PTEN gene wasdetected by in situ immunohistochemistry. Flow cytometry, electron microscope were adopted to measure the effects of PTEN on cell cycle, ultrastructure, and cell growth of the transfected HHCC, as well as on tumorigenicity in nude mice. RESULTS PTEN was expressed in the HHCC transfected with PTEN gene by in situ immunohistochemistry. The progression of cell cycle was arrested from G 1 to S phase. Compared with the parent HHCC cells, the growth of cells transfected with PTEN gene in vitro and in nude mice were dramatically inhibited by 70.6% and 72.2%, respectively. CONCLUSION HHCC hepatoma carcinoma cells could be arrested at G 1/S phase and the growth of cells could be significantly suppressed by exogenous PTEN gene.