hPOT1基因过表达对HeLa细胞细胞周期和凋亡的影响

Effects of hPOT1 overexpression on cell cycle and the apoptosis of HeLa cells

目的观察人POT1(protectionoftelomeres1)基因过表达对HeLa细胞细胞周期和细胞凋亡的影响。方法利用本课题组构建的hPOT1基因真核表达重组质粒pcDNA3-hPOT1,经脂质体介导瞬时转染HeLa细胞;通过RT-PCR和EMSA法(电泳迁移率改变分析)检测外源基因的表达效果,流式细胞术分析细胞周期,Hoechst33342荧光染色检测细胞凋亡。结果pcDNA3-hPOT1重组质粒转染HeLa细胞48h后,mRNA和蛋白质分析表明,外源性hPOT1基因能在HeLa细胞中有效表达,HeLa细胞阻滞于细胞周期S期,而对凋亡无明显影响。结论hPOT1基因可能参与了高等真核细胞细胞周期调控过程,但与细胞凋亡无密切关系。

ObjectiveTo observe the effects of hPOT1 overexpression on cell cycle and apoptosis of HeLa cells. MethodsBy using an eukaryotic expression plasmid of pcDNA-hPOT1, liposomal mediates transient transfection of HeLa cells. The expressing effects of foreign genes were detected by RT-PCR and EMSA method. The cell cycles were analyzed by flow cytometry. And cell apoptosis was detected by Hoechst 33342 fluorescent staining. ResultsAfter 48 hours of the pcDNA3-hPOT1 transfected into HeLa cells, mRNA and protein analysis showed that hPOT1 could be efficiently expressed. Cell cycle in HeLa cells was blocked in S phase, and the overexpression of hPOT1 didnt show the effects on apoptosis. ConclusionThose results proved that hPOT1 functioned in the regulation of cell cycle in eukaryotic cells but not in cell apoptosis.