pEgr-hPTEN体外稳定转染对人脑胶质瘤SHG-44细胞周期及增殖的影响

Effect of pEgr-hPTEN steadily transfected on the cell cycle progression and proliferation of SHG-44 human glioma cells

目的 探讨外源野生型PTEN基因对人脑胶质瘤SHG 4 4细胞周期及增殖的影响 ,阐明PTEN基因抑制肿瘤细胞增殖的机制。方法 脂质体包裹重组质粒pEgr hPTEN体外转染内源性PTEN基因突变的SHG 4 4胶质瘤细胞 ;G4 18筛选稳定表达细胞株并扩增培养 ;采用细胞计数法检测肿瘤细胞增殖速度 ;流式细胞术检测肿瘤细胞周期的变化。结果 经 10到 2 0dG4 18筛选得到稳定表达细胞株 ;重组质粒pEgr hPTEN稳定转染可明显抑制SHG 4 4细胞的体外增殖 ,细胞周期明显改变 ,细胞从G1期到S期发生阻滞。结论 提示转染外源野生型PTEN基因可使SHG 4 4细胞周期阻滞于G1期 ,并可抑制肿瘤细胞增殖。

Objective To investigate the effect of exogenous wild type PTEN gene transfer on the cell cycle progression and proliferation of SHG-44 human glioma cells, and to determine the mechanism that PTEN gene inhibits the proliferation of tumor cells. Methods pEgr-hPTEN recombinant plasmids were packed with liposome to transfect glioma cell line SHG-44, which containing the endogenous mutant PTEN gene. The stable express cells were selected by G418.The proliferation of tumor cells were examined by cell counted and the changes of cell cycle progression were observed by FACscan method. Results After 10 to 20 days G418 selection, stable express cells were obtained. Growth of SHG-44 was inhibited significantly by stable pEgr-hPTEN transfer in vitro. The progression of cell cycle were arrested from G1 to S phase. Conclusion The growth of SHG-44 glioma cells could be arrested at G1 phase, and it could be significantly suppressed by exogenous PTEN gene transfer.