PTEN基因转染对卵巢癌SKOV3细胞生长抑制和诱导凋亡的研究

目的研究外源性PTEN基因转染对人卵巢癌SKOV3细胞生长增殖抑制和凋亡诱导作用,探索卵巢癌的新治疗方法。方法将携带PTEN基因的真核表达载体pBP-PTEN和不含该基因的空载体pBP转染卵巢癌SKOV3细胞,实验分组为pBP-PTEN、pBP和SKOV3组,四甲基偶氮唑蓝(MTT)法观察对SKOV3细胞的生长抑制作用,Annexin V/PI双标记流式细胞术检测转染后对SKOV3细胞的凋亡诱导作用,Annexin V/PI双染荧光显微镜观察细胞凋亡形态。结果MTT检测,0.1、0.2和0.3μg/孔PTEN质粒DNA转染SKOV3细胞48 h,生长抑制率分别为13.31%、27.67%、42.25%,不同时间点(24、48和72 h)检测,细胞生长抑制率存在剂量-时间依赖关系。光镜观察,转染24 h后即可见明显的形态学改变。流式细胞术检测,4μg/孔PTEN质粒DNA转染SKOV3细胞24和48 h后,细胞凋亡率分别达38.08%和65.60%(P<0.01)。Annexin V/PI双染荧光显微镜观察,细胞凋亡率存在时间-剂量以来关系。结论PTEN基因转染抑制人卵巢癌细胞增殖并显著诱导细胞凋亡。

Identification of inhibitory effect on proliferation and induction of apoptosis in human ovarian carcinoma SKOV3 cells by wild-type PTEN gene transfection

Objective The effects of proliferation inhibition and apoptotic induction in human ovarian carcinoma SKOV3 cells,by means of wild-type PTEN transfection,were identified,in an attempt to develop a novel therapy for human ovarian cancer.Methods Eukaryotic expression vectors containing PTEN gene or without PTEN gene were transfected into human ovarian carcinoma SKOV3 cells.The inhibition of cell proliferation was determined by MTT in vitro.Induction of apoptosis was identified by flow cytometry,using an Annexin V/PI kit.Moreover,apoptotic evidence was investigated by both microscopy and fluorescence microscopy after staining by the Annexin V/PI kit.Results The growth and proliferation ratios in the cells were 13.31%,27.63% and 42.28%,respectively,after 0.1,0.2 and 0.3 μg/well transfection with the plasmid containing wild-type PTEN for 48 h.The proliferation inhibition was investigated after transfected with PTEN at the different concentrations of the plasmid for 24,48 and 72 h,for further identifying the inhibitory effects.The cells were inhibited by PTEN,in a dose-time-dependent manner.The apoptotic cells analyzed by flow cytometry accounted for 38.08% and 65.60%,after transfected with 4 μg/well PTEN plasmid for 24 and 48 hours,respectively.The results come from fluorescence microscopy were similar to that from flow cytometry.After transfected with PTEN for 24 h,the apoptotic morphological changes emerged under a microscope.Conclusion PTEN gene transfection obviously inhibited cell growth and proliferation and induced apoptosis in human ovarian carcinoma SKOV3 cells in vitro.