17β-雌二醇对人子宫内膜癌细胞的增殖及P21ras和p-Erk表达的研究

Effect of 17β-estradiol on proliferation and expressions of P21ras and p-Erk in endometrial carcinoma cells

目的:探讨17β-雌二醇(E2)对人子宫内膜腺癌雌激素受体(ER)阳性的Ishikawa和ER阴性的HEC-1A细胞增殖,细胞周期及其对丝裂原活化蛋白激酶(MAPK)通路相关蛋白P21ras和p-Erk表达的影响及意义。方法:用MTT法,流式细胞技术检测不同浓度E2作用Ishikawa和HEC-1A细胞不同时间的细胞吸光度值及细胞周期,用免疫细胞化学法检测上述细胞中P21ras和p-Erk的表达。结果:随E2浓度增加,Ishikawa细胞吸光度值上升并呈时间依赖性(P<0.01),G0～G1期比例下降(P<0.01),S期比例升高(P<0.01);HEC-1A细胞吸光度值及细胞周期无明显变化(P>0.05);10-6mol/LE2作用于Ishikawa细胞30min时,P21ras和p-Erk活化表达最强,作用于HEC-1A细胞15min时,P21ras和p-Erk即有明显表达而且达高峰,随着E2浓度增加两种细胞中P21ras和p-Erk表达均逐渐增加,呈浓度依赖性。结论:E2可促进子宫内膜癌Ishikawa细胞的增殖和周期进展,而且可以促进Ishikawa和HEC-1A中P21ras和p-Erk表达增加。

Objective：To explore the influence of β-estradiol E2 on proliferation, cell cycle progression, expressions of P21ras and p-Erk in endometrial carcinoma cells Ishikawa and HEC-1A. Methods：E2 was used to stimulate Ishikawa and HEC-1A cells, the proliferation and cell cycle of Ishikawa and HEC-1A were detection by MTr and fluorescence-activated cell sorting technique. The expression of P21ras and p-Erk were examined by immunoprecipitation in Ishikawa and HEC-1A cell after stimulated with 106mol/L E2 for different times. Results： With increased concentrations of E2, the proliferation of Ishikawa cell increased gradually and in a time-dependent manner（ P 〈 0.01 ） , percentage of Ishikawa cell at GO - G1 phase decreased （ P 〈 0.01 ） and percentage of S phase cells increased significantly （ P 〈 0.01 ）, whereas that of HEC-1A cell did not show significant alteration. The maximal activation of P21ras and p-Erk took place at 30min in Ishikawa cell and 15min in HEC-1A cell after stimulation with 10^- 6 moL/L E2 in HEC-1A cell. With increased doses of E2 ,the activation of P21ras and p-Erk increased gradually and presented in concentration-dependent manner. Conclusion： 17β-estradiol not only promotes the cell proliferation and the cell cycle distrubion of endometrial carcinoma cell lines, but also activates promptly P21ras and p-Erk expression in endometrial carcinoma cells.