孕激素对孕激素受体阴性的子宫内膜腺癌细胞JEC的影响

Effects of Progesterone on Growth of Endometrial Adenocarcinoma Cell Line JEC with Negative Progesterone Receptor Negative

目的探讨不同浓度的孕激素(progesterone,P)对孕激素受体(progesterone receptor,PR)阴性的子宫内膜腺癌细胞系JEC细胞增殖和细胞周期的影响及其机制。方法取JEC细胞体外培养,加入含不同浓度P的培养液,采用细胞计数法、四甲基偶氮唑蓝(MTT)比色法和流式细胞术(FCM)的方法,观察JEC细胞的增殖活性和细胞周期时相变化;同时用FCM检测加入P前后JEC细胞中p21蛋白的表达;用免疫组织化学法及图像分析,检测加入P前后JEC细胞内Cyclin A的表达。结果 (1)细胞计数法和MTT法显示,P可使JEC细胞数明显减少;(2)1×10-5 mol/L的P作用72 h,可使JEC细胞G0/G1期比例增加,S期及G2/M比例减少;(3)P对JEC细胞内p21蛋白的表达无影响。(4)P可明显抑制JEC细胞内Cyclin A蛋白的表达。结论 PR阴性的JEC细胞生长可受到孕激素的调控,其作用具有剂量和时间效应.这种调控可能与Cyclin A有关,但与P21蛋白无关。

Objective To study the effect of different concentration of progesterone（P） on the growth and cell cycle of endometrial adenocarcinoma cell line JEC with negative progesterone receptor in vitro and its possible mechanism.Methods Two groups were set up：sample group（progesterone at different concentrations） and control group without progesterone.The proliferative capacity of endometrial adenocarcinoma cell line JEC in the culture medium with progesterone was assessed by cell counting on a haemocytomter and evaluated by the means of 3-（4,5-dimethylthiazol-z-yl）-2,5-dipheny tertrazolium blue（MTT） and cell cycle was analyzed by flow cytometry（FCM）.The expression of p21 in JEC was examined by FCM,and the expression of Cyclin A of JEC was examined by immunocytochemical staining and using automatic image analysis technology.Results（1）The cell counting and MTT showed that the cell numbers had significant difference in the other experimental groups compared with the control group.（2）FCM showed that progesterone（10-5mol/L） increased the cells percentage at the G0/G1 phases and decreased the cells percentage in S and G2/M phases.（3）The expression of P21 protein have no change after dealing with progesterone.（4）The expression of Cyclin A could be inhinbited obviously by progesterone.Conclusion It is suggested in the present study that progesterone inhibits the proliferation of endometrial adenocarcinoma cell line JEC in vitro which is in dose— and time—dependent manner.Its effect may be associated with the change in the expression of Cyclin A,butuncorrelated with P21 protein.