孕三烯酮对体外培养异位子宫内膜细胞生长及凋亡的影响

目的　探讨孕三烯酮对体外培养的子宫内膜异位症(内异症)患者的异位子宫内膜细胞(异位内膜细胞)生长及凋亡的影响,及第10号染色体缺失的磷酸酶和张力蛋白同源物(phosphataseandtensionhomologuedeletedonchromosome10,PTEN)基因在异位内膜细胞表达的变化。方法　以0、10. 6和10 .4 mol/L浓度的孕三烯酮,对体外培养的异位内膜细胞分别进行处理;采用四甲基偶氮唑蓝比色法,检测孕三烯酮对异位内膜细胞作用0 ~48h的细胞抑制情况并绘制细胞生长曲线;应用透射电镜观察孕三烯酮对异位内膜细胞作用24h时的细胞超微结构变化;采用流式细胞仪检测异位内膜细胞凋亡率及细胞周期变化;应用PTEN单克隆抗体,经流式细胞仪分析异位内膜细胞PTEN基因的表达。结果　孕三烯酮作用于异位内膜细胞8、16、24、32、40和48h的细胞生长率,在孕三烯酮浓度为10 .6 mol/L时,分别为99 .6%、87 .3%、79 .8%、62 .3%、51. 7%和44. 2%;在10 .4 mol/L时,分别为99 .2%、77 .1%、69. 6%、51 .1%、33 .7%和23 .6%。同一浓度各作用时间比较,差异均有统计学意义(P<0 .05)。孕三烯酮作用24h后,异位内膜细胞发生典型的细胞凋亡形态学改变。浓度为10 6、10 4 mol/L的孕三烯酮作用于异位内膜细胞后,凋亡率分别为1 3%、15 .0%;浓度为0mol/L的孕三烯酮

Effect of gestrinone on growth and apoptosis in isolated ectopic endometrium cells in vitro

Objective To investigate the effects of gestrinone on growth and apoptosis, as well as the expression of phosphatase and tension homologue deleted on chromosome 10(PTEN) in isolated ectopic endometrium cells in vitro and the underlying mechanisms. Methods Ectopic endometrium cells were cultured and exposed to gestrinone of different doses of 0, 10 -6 and 10 -4 mol/L respectively. The inhibition of the cells during 48 hours was determined by methylthiazolyl tetrazolium (MTT) assay, and the cell growth curve was made. Gestrinone was administered to the cells and at 24 hours the morphological changes were observed by transmission electron microscopy and the apoptosis rate, cell cycle and PTEN expression were monitored by flow cytometry (FCM) at the same time. Results Gestrinone at different concentrations could inhibit the growth and proliferation of ectopic endometrium cells in a dose- and time-dependent manner. The inhibition rate of cell growth after exposed to gestrinone for 8,16,24,32,40 and 48 h was 99.6%,87.3%,79.8%,62.3%,51.7% and 44.2% in the 10 -6 mol/L group,and 99.2%,77.1%,69.6%,51.1%,33.7% and 23.6% in the 10 -4 mol/L group (P<0.05),and cell growth curve was changed accordingly. After 24 hour exposure to gestrinone from 10 -6 to 10 -4 mol/l, apoptotic changes of cells were observed under transmission electron microscope. FCM showed that after the exposure to gestrinone, the apoptotic rate of ectopic endometrium cells was 1.3% in 10 -6 mol/L group and 15.0% in 104 mol/L group. It was significantly increased when compared with the 0 mol/L group,the apoptotic rate of which was 0% (P<0.05). The level of PTEN expression of the ectopic endometrium cells was 60.6% after treated with 0 mol/L gestrinone, while in 10 -6 and 10 -4 mol/l groups the level of PTEN expression was increased to 75.3% and 85.7%, significantly higher than that of the 0 mol/L group (P<0.05). Conclusion Gestrinone can significantly inhibit the growth and proliferation of ectopic endometrium cells, and this effect was related to increase of PTEN expression.