GPER在雌激素激活的子宫内膜癌细胞内PI3K／Akt信号通路中的作用  

Effect of GPER on the activation of PI3K/Akt induced by 17β-estradiol in endometrial carcinoma cells

作　　者：张燕彩[1] 郭瑞霞[1] 葛新[1] 乔玉环[1] 

机构地区：[1]郑州大学第一附属医院妇产科,450052

出　　处：《中华妇产科杂志》2012年第4期292-296,共5页Chinese Journal of Obstetrics and Gynecology

基　　金：国家自然科学基金（81072124）

摘　　要：目的探讨G蛋白偶联ER（GPER）在1713雌二醇（1713-E2）激活的子宫内膜癌细胞内磷脂酰肌醇3激酶（P13K）／蛋白激酶B（Akt）信号通路中的作用。方法应用免疫组化sP法检测子宫内膜癌细胞系HEC-1A和Ishikawa细胞中GPER、ERa、ERl3、Akt和磷酸化Akt（p-Akt）蛋白的表达部位。应用蛋白印迹法检测1×10μmol／L的17β-E2，处理不同时间（分别为0、15、30、60、120min）后HEC-1A和Ishikawa细胞中Akt蛋白活化水平及GPER、ERa和ER[3蛋白表达水平的变化。结果免疫组化SP法检测显示，在HEC-1A细胞中，GPER、Akt、p-Akt蛋白在细胞质呈棕黄色阳性表达，ERa蛋白在细胞核呈棕黄色阳性表达，ER13蛋白呈阴性表达；在Ishikawa细胞中，GPER、Akt、p-Akt蛋白在细胞质中均呈棕黄色阳性表达，ERa、ER[3蛋白在细胞核呈棕黄色阳性表达。蛋白印迹法检测显示，1×10μmol／L的1713-E2处理后，Ishikawa和HEC—1A细胞中GPER蛋白的表达水平从处理15min开始即明显升高（P〈0．05），其中Ishikawa细胞在处理30min时达最高（为0．192±0．004），HEC-1A细胞在处理15min时达最高（为0．184±0．006）；lshikawa和HEC-1A细胞中Akt蛋白的活化水平从处理15min开始即明显升高（P〈0．05），其中Ishikawa细胞在处理30min时达最高（为0．666±0．021），HEC-1A细胞在处理15min时达最高（0．7884-0．035）；而Ishikawa和HEC．1A细胞中ERd和ER13蛋白的表达水平无明显变化（P〉0．05）。结论GPER可能介导了P13K／Akt信号通路的活化，参与了子宫内膜癌的非基因转录效应。Objective To investigate the expression of G protein-coupled ER （GPER） and ER in the activation of phosphatidylinositol 3-kinase （ PI3K ）/protein kinase B （ Akt ） induced by 17 β-estradiol （1713-E2） in endometrial carcinoma cells, Ishikawa and HEC-1A. Methods Expressions of GPER, ERa and ERβ protein in Ishikawa and HEC-1A cells were detected by immunohistochemical SP method. Levels of GPER, ERa and ERβ were examined by western blot in Ishikawa and HEC-1A cells after treated with 1 x 10 6 mol/L 17β-E2 at different time （0,15,30,60,120 minutes）. Results GPER was positive expressed in Ishikawa and HEC-1A cells. ERct and ERβ were both positive expressed in Ishikawa cells. While, ERec was weakly expressed and ER/3 was almost negatively expressed in HEC-1A cells. Western blot analysis showed that 1 x 10-6 moL/L 17~_E2 treatment, the Ishikawa and HEC-1A cells GPER protein level for 15 minutes markedly increased （P 〈 0. 05 ）, which Ishikawa 30 minutes, when cells reached the highest level （0. 192 0. 004） , HEC-1A cells for 15 minutes and reached the highest level （0. 184 -＋0. 006） ; Ishikawa and HEC- 1A cells, Akt, activation of 15 minutes from the treatment start was significantly increased （P 〈 0. 05 ） , which Ishikawa cells for 30 minutes and reached the highest level （0. 666 ＋ 0. 021 ） , HEC-1A cells for 15 minutes and reached maximum （0. 788 ~ 0. 035）; Ishikawa and HEC-IA cells, ERcc and ERI3 protein expression did not change significantly （ P 〉 0. 05 ）. Conclusion GPER likely involved in non-nuclear activation of PI3K/Akt signaling pathways in endometrial carcinoma cells, Ishikawa and HEC-1A.

关 键 词：子宫内膜肿瘤 受体 雌激素 受体 G-蛋白偶联 雌二醇 1-磷脂酰肌醇3-激酶 原癌基因蛋白质c—akt 

分 类 号：R737.33[医药卫生—肿瘤]