PTEN基因参与介导孕三烯酮诱导的异位子宫内膜细胞凋亡.docVIP

PTEN基因参与介导孕三烯酮诱导的异位子宫内膜细胞凋亡 【关键词】 PTEN基因 【摘要】 目的 探讨孕三烯酮对子宫内膜异位症细胞中PTEN基因表达的影响及对细胞凋亡的诱导作用。方法 体外原代培养人子宫内膜异位症细胞，经不同浓度孕三烯酮作用后，采用四甲基偶氮唑兰（MTT）比色法、透射电镜、流式细胞仪和核DNA琼脂糖凝胶电泳检测细胞的凋亡情况，应用Western免疫印记法，观察药物作用前后PTEN基因的表达变化。结果 原代培养的子宫内膜异位症细胞经孕三烯酮处理后，细胞生长被明显抑制，并呈剂量-时间依赖性;透射电镜下可见细胞核染色质浓缩边集、胞浆浓缩、核碎裂及凋亡小体形成等凋亡的典型形态学变化;流式细胞仪检测提示，细胞周期从G 1 期到S期发生抑制，并在G 1 峰前出现明显的凋亡峰（15.0%）;细胞核DNA琼脂糖凝胶电泳呈现凋亡细胞特有的梯状条带;PTEN基因的表达显著上调（P0.05）。未经孕三烯酮作用的子宫内膜异位症细胞无明显凋亡表现，PTEN基因呈低水平表达。结论 孕三烯酮能够诱导子宫内膜异位症细胞的凋亡并上调PTEN基因的表达，这可能是孕三烯酮治疗子宫内膜异位症的分子机制之一。 关键词 子宫内膜异位症 抑癌基因 PTEN 孕三烯酮 凋亡 【文献标识码】 A 【文章编号】 1606-8106（2004）15-1547-03 PTEN gene is involved in gestrinone induced apoptosis of ectopic endometrium cells Ma Jiajia，Chen Biliang，Ma Xiangdong，et al. Department of Gynecology and Obstetrics，Xijing Hospital of the Fourth Military Medical University，Xi’an710033. 【Abstract】 Objective To find out the effects of gestrinone on the expression of PTEN gene and to investigate the underlying mechanism of inhibition and apoptotic action that caused by gestrinone on isolated ectopic endometrium cells in vitro.Methods Ectopic endometrium cells were cultured in vitro.After being cultured with different dose of gestrinone for different time，the cells were examined by MTT assay，transmission electronic microscope，flow cytometry and nucleus DNA agarose gel electrophoresis to detect the apoptotic changes in them.The changes of PTEN gene exˉpression were detected by western-blot analysis.Results Gestrinone at different concentrations can inhibit the growth and proliferation of ectopic endometrium cells in a dose-and time-dependent manner.The typical changes of ultrastructure included peripheral accumulation of nuclear chromation，fragmentation of nuclear and apoptosis body.Cell cycle determined by flow cytometry showed that the percentage of G 1 phase increased，but the percentage of S phase decreased;one visible apoptosis peak（15.0%）in front of the G 1 phase was also found.The characteristic DNA ladder was found in the agarose gel electropho