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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 .
Objective To study the effect of exogenous PTEN gene on the cell cycle, proliferation and tumorigenicity of human hepatocellular carcinoma cell line HHCC and to explore a new way of gene therapy for hepatocellular carcinoma.Methods The eukaryotic expression vector carrying PTEN gene was transfected into HHCC cells in vitro , And stably transfected cell clones were screened and expanded for culture. The expression of PTEN gene was detected by immunohistochemistry. The expression of PTEN gene was detected by flow cytometry, electron microscopy and growth curve analysis. The cell cycle, ultrastructure , Growth rate and other characteristics, and observe the effect of human PTEN gene on the ability of tumorigenesis of HHCC nude mice.Results The stable expression of PTEN protein in HHCC hepatoma cells stably transfected with PTEN gene, the cell cycle significantly changed from G1 phase S-phase inhibition, the cell ultrastructure degenerative changes, cell proliferation activity decreased by 70.6% nude mice tumorigenic ability inhibition rate 72.2% .Conclusion Transfection of exogenous PTEN gene can prevent HHCC hepatocellular carcinoma cells from the G1 S phase into the period, and inhibit tumor cell proliferation.