睾酮在乳腺癌细胞中对FEN1表达的影响及其机制-第三军医大学学报.doc

睾酮对乳腺癌细胞中FEN1表达的影响 王 杨，陈 彬，姜晓梅，王 越，付晓红，何凤田（400038重庆，第三军医大学生物化学与分子生物学教研室） [摘要]目的 探讨睾酮（testerone，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（letrozole）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 与对照组相比，加入雌二醇后MCF-7细胞中FEN1 mRNA的表达升高2倍（P＜0.01），当加入ICI和U0126后分别降低10.6倍和2.2倍（P＜0.01）。加入睾酮后MCF-7aro细胞中FEN1 mRNA的表达升高1.7倍（P＜0.01），蛋白的表达升高1.8倍（P＜0.01）；当加入来曲唑后FEN1 mRNA的表达下降2.4倍，蛋白的表达降低1.8倍，当加入U0126后蛋白的表达降低2.3倍（P＜0.01）；加入睾酮后ERK和Elk-1的磷酸化水平分别升高2.3倍和2.6倍（P＜0.01），当加入来曲唑后ERK和Elk-1的磷酸化水平分别降低2.6倍和2.4倍（P＜0.01），当加入U0126后ERK和Elk-1的磷酸化水平分别降低10.4倍和148.8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。 [关键词]乳腺癌；睾酮；FEN1 中图分类号：R737. 31 文献标识码：A Effect and mechanism of testerone on the expression of FEN1 in breast cancer cells Wang Yang， Chen Bin， Jian Xiao-Mei， Wang Yue， Fu Xiao-Hong， He Fengtian （Department of Biochemistry and Molecular Biology ， Third Military Medical University， Chongqing， 400038， China） [Abstract] Objective To explore the effect and mechanism of testerone on the expression of FEN1 in breast cancer. Methods The expression of FEN1 in MCF-7 cells was observed by RT-PCR when cells were treated with 17β-estradiol（E2） or E2 combined with ICI182，780， an estrogen receptor antagonist or E2 combined with U0126，a MEK inhibitor. In MCF-7 cells over-expressing aromatase （MCF-7/Aro）， the expression of FEN1 was observed by RT-PCR when cells 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 blot when cells were treated with testerone or testerone combined with letrozole or testerone combined with U0126. Results In MCF-7 cells ,E2 could increase the expression of FEN1 mRNA by 2 folds（P＜0.01），which could be inhibited by ICI182，780 o