抑癌基因PTEN真核表达载体的构建及对人卵巢癌细胞系SW626增殖的影响

目的 构建抑癌基因PTEN的真核表达载体,研究外源性PTEN基因稳定表达对人卵巢癌细胞系SW626体外增殖的影响.方法 构建PTEN基因的真核表达载体pcDNA3.1/PTEN,转染重组质粒pcDNA3.1/PTEN于体外培养的人卵巢癌细胞系SW626,筛选并获得稳定表达PTEN基因的细胞克隆,应用RT-PCR和western blot分析PTEN基因的mRNA及蛋白质表达,MTT实验分析细胞增殖能力.结果 稳定转染PTEN基因的细胞系有外源目的 基因的mRNA及其蛋白表达.MTT实验表明,pcDNA3.1/PTEN 转染组细胞增殖能力低于pcDNA3.1(-)转染组和对照组 (P＜0.05).结论 外源性抑癌基因PTEN的稳定表达可抑制人卵巢癌细胞系SW626的增殖.

Construction of PTEN Eukaryotic Expression Plasmid and Its Effect on Human Ovarian Cancer Cell Line SW626

Objective To construct PTEN eukaryotic expression plasmid and investigate the effect of exogenous wild PTEN gene stably transfection on growth of human ovarian cancer cell line SW626 in vitro.Methods Recombinant pcDNA3.1/PTEN plasmid was constructed and transfected into the SW626 cell line by lipofectamine 2000.Cell clones stably expressing PTEN were obtained by detecting the mRNA and protein expression of PTEN in G418 resistant clones.Cell proliferation was tested by MTT assay.Results The PTEN stably transfected cells demonstrated the PTEN mRNA and protein expression.There was a significant decline in cell proliferation of pcDNA3.1/PTEN transfected SW626 cells in comparison with the mock transfected ones(P<0.05).Conclusion Stable expression of exogenous PTEN can suppress the growth of human ovarian cancer cell line SW626.