GPR30下调对子宫内膜癌细胞及裸鼠移植瘤组织PI3K/Akt信号通路的影响  

Influence of down-regulation of GPR30 on PI3K/Akt signaling pathway in endometrial carcinoma cells and tumor tissue of nude mice

作　　者：雷冬梅[1] 郭瑞霞[2] 刘泇希 葛新[2] 乔玉环[2] 

机构地区：[1]郑州大学第三附属医院病理科,郑州450052 [2]郑州大学第一附属医院妇产科,郑州450052

出　　处：《郑州大学学报（医学版）》2015年第5期593-596,共4页Journal of Zhengzhou University(Medical Sciences)

基　　金：国家自然科学基金项目81072124

摘　　要：目的:探讨G蛋白偶联受体(GPR30)下调对子宫内膜癌细胞及裸鼠移植瘤组织PI3K/Akt信号通路的影响。方法:采用免疫细胞化学SP法观察子宫内膜癌细胞系HEC-1A和Ishikawa细胞中GPR30、Akt和磷酸化Akt(p-Akt)蛋白的定位。以p GFP-V-RS(对照)和p GFP-V-RS-GPR30(干扰)分别转染两种细胞,用蛋白印迹法检测两种细胞中GPR30、Akt及p-Akt蛋白的表达水平。构建裸鼠移植瘤模型(分别接种上述4种转染细胞,共4组),用免疫组化SP法检测4组裸鼠移植瘤组织中p-Akt蛋白的表达。结果:1HEC-1A和Ishikawa细胞中,GPR30、Akt和p-Akt蛋白阳性表达均呈棕黄色,定位于细胞质。2稳定转染p GFP-V-RS-GPR30的两种细胞株中GPR30和p-Akt的表达均下调(P<0.05)。3与接种转染p GFP-V-RS细胞的2组裸鼠相比,接种转染p GFP-V-RS-GPR30细胞的2组裸鼠移植瘤组织中p-Akt表达降低。结论:在子宫内膜癌细胞及裸鼠移植瘤组织中,GPR30下调可能抑制了PI3K/Akt通路的活化。Aim：To investigate the influence of down-regulation of G protein-coupled estrogen receptor 30 （ GPR30 ） on activation of PI3K/Akt signaling pathway in endometrial carcinoma cells and tumor tissue of nude mice .Methods：The location of GPR30, Akt and p-Akt protein in HEC-1A and Ishikawa cells was detected by immunocytochemical SP method . The expression of GPR30 in HEC-1A and Ishikawa cells was down-regulated by transfection with pGFP-V-RS-GPR30, a GPR30 antisense expression vector , and the cells transfected with pGFP-V-RS were the control; the levels of GPR30,Akt and p-Akt were detected by Western blot .The nude mice were allocated into 4 groups and inoculated the above transfected cells, and immunohistochemistry was performed to observe the changes of the expression level of p -Akt in the xenograft tis-sue.Results：Immunocytochemical SP method showed that GPR 30, Akt and p-Akt was stained as brown and yellow in cell cytoplasm of HEC-1A and Ishikawa cells.Western blot analysis showed that the expressions of GPR 30 and p-Akt were de-creased in tumor cells transfected by pGFP-V-RS-GPR30 compared with the control （P〈0.05）.The expression of p-Akt was decreased in the mice inoculated the tumor cells transfected by pGFP-V-RS-GPR30 .Conclusion： Down-regulation of GPR30 inhibit the activation of PI3K/Akt signaling pathway in Ishikawa and HEC-1A cells and tumor tissue of nude mice .

关 键 词：子宫内膜肿瘤 受体 G蛋白偶联 磷脂酰肌醇3激酶 HEC-1A细胞 ISHIKAWA细胞 裸鼠 

分 类 号：R737.33[医药卫生—肿瘤]