睾酮对乳腺癌细胞中FEN1表达的影响  

Effect of testosterone on FEN1 expression in breast cancer cells

作　　者：王杨[1] 陈彬[1] 姜晓梅[1] 王越[1] 付晓红[1] 何凤田[1] 

机构地区：[1]第三军医大学基础医学部生物化学与分子生物学教研室,重庆400038

出　　处：《第三军医大学学报》2013年第2期95-98,共4页Journal of Third Military Medical University

基　　金：国家自然科学基金(81272908);重庆市自然科学基金(CSTC2012jjA10121);第三军医大学归国人员启动基金(2009)~~

摘　　要：目的探讨睾酮(testosterone,T)在乳腺癌细胞中对瓣状核酸内切酶(flap endonuclease 1,FEN1)表达的影响及其机制。方法以MCF-7作为研究对象,采用RT-PCR观察雌二醇(17β-estradiol,E2)单独作用以及分别与雌激素受体(estrogen receptor,ER)拮抗剂ICI182,780(ICI)和MAPK途径抑制剂U0126共同作用时FEN1表达的变化。以MCF-7aro(芳香化酶过表达的MCF-7)作为研究对象,采用RT-PCR观察加入单独的睾酮以及睾酮和芳香化酶抑制剂来曲唑(letro-zole)共同作用时FEN1表达的变化;采用Western blot观察睾酮单独作用以及分别与来曲唑和U0126共同作用时FEN1、p-ERK和p-Elk的变化。结果与对照组比较,加入雌二醇后MCF-7细胞中FEN1 mRNA的表达升高2.04倍(P<0.01),加入ICI和U0126后分别降低10.63倍和2.17倍(P<0.01)。加入睾酮后MCF-7aro细胞中FEN1 mRNA的表达升高1.66倍(P<0.01),蛋白的表达升高1.80倍(P<0.01);加入来曲唑后FEN1 mRNA的表达下降2.38倍,蛋白的表达降低1.84倍,加入U0126后蛋白的表达降低2.28倍(P<0.01);加入睾酮后ERK和Elk-1的磷酸化水平分别升高2.28倍和2.60倍(P<0.01),加入来曲唑后ERK和Elk-1的磷酸化水平分别降低2.60倍和2.37倍(P<0.01),加入U0126后ERK和Elk-1的磷酸化水平分别降低10.38倍和119.50倍(P<0.01)。结论睾酮可通过MAPK途径上调MCF-7 aro细胞中FEN1的表达。Objective To determine the effect of testosterone on the expression of flap endonuclease 1 （ FEN1 ） in breast cancer. Methods The expression of FEN1 in MCF-7 ceils was observed by RT-PCR after the ceils were treated with 17β-estradiol （E2）, E2 combined with ICI182, 780, an estrogen receptor antago- nist, and E2 combined with U0126, a MEK inhibitor, respectively. In MCF-7 ceils over-expressing aromatase （MCF-7/Aro）, the expression of FEN1 was observed by RT-PCR when ceils were treated with testerone or testerone combined with letrozole, one of aromatase inhibitors. The expression of FEN1 protein, p-ERK and p-Elk was observed by Western blotting when the cells were treated with testosterone, testosterone combined with letrozole, or combined with U0126. Results In MCF-7 cells, E2 resulted in an increase in the expression of FEN1 mRNA by 2.04 folds （P 〈0.01 ）, which was inhibited by ICI182,780 or U0126 （ 10.63 and 2. 17 folds, P 〈0. 01 ）. In MCF-7aro ceils, testosterone resulted in an increase in the expression of FEN1 at mRNA and protein levels by 1.66 and 1.80 folds respectively（P 〈 0.01 ）, which was inhibited by letrozole （2.38 and 1.84 folds, P 〈0. 01 ） or U0126（2.28 folds for protein only, P 〈0. 01 ）. Testosterone increased the phosphory- lation of p-ERK and p-Elk by 2.28 and 2.60 folds, which was inhibited by letrozole by 10.38 folds or U0126 by 119.50 folds（P 〈0.01）. Conclusion Testosterone up-regulates FEN1 in MCF-7aro cells through MAPK signaling pathway.

关 键 词：乳腺肿瘤 细胞系 肿瘤 睾酮 DNA瓣核酸内切酶类 

分 类 号：R737.9[医药卫生—肿瘤] R968.1[医药卫生—临床医学]