逆转录病毒介导PTEN基因抑制人肝癌细胞系的生长

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。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。

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

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.