外源性子宫珠蛋白基因对宫颈癌HeLa细胞增殖及凋亡的影响

Effects of exogenous uteroglobin gene on cell apoptosis and proliferation of human cervical cancer cell line HeLa

目的探讨人子宫珠蛋白(UG)基因对宫颈癌Hela细胞系细胞增殖及凋亡的影响。方法构建真核细胞表达载体pcDNA3.1-UG(+),以脂质体介导法稳定转染体外培养的人宫颈癌Hela细胞,同时转染空载体pcDNA3.1质粒,以未转染细胞为对照,转染成功后分别命名为pcDNA3.1-UG(+)/Hela组、pcDNA3.1/Hela组、Hela组。应用逆转录聚合酶链式反应(RT-PCR)、Westernblot方法分析目的基因表达,并采用四甲基偶氮唑蓝(MTT)法和流式细胞术检测细胞增殖和细胞凋亡。结果pcDNA3.1-UG(+)/Hela组UGmRNA及UG蛋白出现明显的高表达,与对照组细胞相比差异有统计学意义(P<0.05);pcDNA3.1-UG(+)/Hela组细胞生长速度明显慢于未转染Hela细胞,其细胞凋亡率与未转染Hela细胞相比差异也有统计学意义;然而pcDNA3.1/Hela组细胞生长速度和凋亡率均无明显变化。结论转染UG基因能诱导Hela细胞凋亡及抑制Hela细胞生长。

Objective To investigate the effects of exogenous wild uteroglobin （UG） gene stable transfection on cell apoptosis and proliferation in human cervical cancer cell line HeLa. Methods UG recombinant eukaryotic expression vector pcDNA3.1-UG （＋） was constructed, and then stably transfected into cultured human cervical cancer HeLa cells by liposome mediation （pcDNA3.1-UG（＋）/HeLa group）. Simultaneously, empty vector pcDNA3.1 plasmid was transfected （pcDNA3.1/HeLa group）. Non-transfected HeLa cells were considered as controls （HeLa group）. RT-PCR and Western blotting assay were applied to detecting the expression of targeted gene and protein, and MTT reduction assay and flow cytometry were applied to measuring cell proliferation and apoptosis. Results UG mRNA and protein were high expressed in the stably transfected cells （pcDNA3.1-UG （＋）/HeLa group）, signifi- cantly higher than those of control group （P〈0.05）. The proliferation rate of cells transfected with pcDNA3.1-UG （＋） plasmid was obviously lower as compared with that of cells transfected with empty vector. The apoptotic rate of pcDNA3.1-UG（＋）/HeLa cells was markedly higher than that of non-transfected HeLa cells. By contrast, there were no significant changes of the cell growth velocity and apoptotic rate in pcDNA3. 1/HeLa group. Conclusion The transfection of UG gene might suppress the growth and induces the apoptosis of human cervical cancer cell line HeLa cells.