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目的 研究抑癌基因PTEN在孕三烯酮诱导体外培养的子宫内膜异位症细胞凋亡中的作用。方法 以不同浓度的孕三烯酮对体外培养人子宫内膜异位症细胞进行处理 ,MTT法测算孕三烯酮作用 0～ 4 8h对子宫内膜异位症细胞的抑制情况并绘制生长曲线 ;取 2 4h为作用时间 ,透射电镜观察细胞超微结构变化 ,流式细胞仪检测细胞凋亡率及细胞周期 ,应用PTEN单克隆抗体经由流式细胞仪分析细胞内PTEN的表达。结果 细胞生长曲线显示孕三烯酮对子宫内膜异位症细胞的生长增殖有显著的抑制作用 ,且呈时间 剂量 效应关系 ;透射电镜观察发现 ,孕三烯酮可诱导子宫内膜异位症细胞发生典型的凋亡形态学改变 ;流式细胞仪检测显示 1× 1 0 -6mol/L和1× 1 0 -4mol/L浓度的孕三烯酮作用 2 4h后 ,子宫内膜异位症细胞的凋亡率分别为 1 3%和 1 5 0 % ,与对照组相比差异显著 ;同时细胞内PTEN的表达有不同程度的上调。结论 孕三烯酮能明显抑制子宫内膜异位症细胞的生长增殖 ,这一作用可能与上调PTEN的表达及诱导细胞凋亡有关
Objective To study the role of tumor suppressor gene PTEN in the induction of apoptosis of endometriosis cells in vitro by gestagentione. Methods Human endometriosis cells were cultured in vitro with different concentrations of gestrinone. The inhibitory effect of gestrinone on the endometriosis cells was measured by MTT assay and the growth curve was drawn The cell ultrastructure was observed by transmission electron microscope (TEM). The apoptosis rate and cell cycle were detected by flow cytometry. The expression of PTEN was analyzed by flow cytometry using PTEN monoclonal antibody. Results The cell growth curve showed that gestrinone inhibited the growth and proliferation of endometriosis cells in a dose-dependent manner. Transmission electron microscopy revealed that gestrinone could induce endometriosis The morphological changes of typical apoptotic cells were observed by flow cytometry. The results of flow cytometry showed that gemtrinone at concentrations of 1 × 10 -6 mol / L and 1 × 10 -4 mol / L, The apoptosis rate of cells was 13% and 150%, respectively, which was significantly different from that of the control group. Meanwhile, the expression of PTEN was up-regulated in different degree. Conclusion Gestrinone can significantly inhibit the growth and proliferation of endometriosis cells, which may be related to the up-regulation of PTEN expression and induction of apoptosis