genistein对人卵巢癌细胞系SKOV_3及其裸鼠移植瘤中表皮生长因子受体介导的信号转导通路的影响  

The effects of genistein on epidermal growth factor receptor mediated signal transduction pathway in human ovarian carcinoma cells lines SKOV_3 and its xenograft in nude mice

作　　者：李昱[1] 米粲[1] 吴永忠[1] 杨素芬[1] 杨正钦[1] 

机构地区：[1]重庆医科大学病理学教研室,400016

出　　处：《中华病理学杂志》2004年第6期546-549,共4页Chinese Journal of Pathology

摘　　要：目的　通过 genistein对人卵巢癌细胞系SKOV3 及其裸鼠移植瘤中表皮生长因子受体(EGFR)介导的肿瘤信号转导系统的影响 ,探讨其抑制增殖作用的机制。方法　应用免疫细胞化学链霉素抗生物素蛋白过氧化物酶 (SP)法检测c erbB 2蛋白的表达 ;Western印迹检测细胞c jun和c fos蛋白的表达 ;逆转录 聚合酶链反应 (RT PCR)检测c erbB 2 ,c raf 1,c jun和c fosmRNA的表达。结果　 2 0 μmol/Lgenistein处理组c erbB 2、c raf 1及其下游基因c jun、c fosmRNA表达减弱。2 0 μmol/Lgenistein处理SKOV3 细胞 4 8h后 ,c erbB 2蛋白表达减弱 ,平均吸光度 (A)值减低 ,为0 4 2± 0 0 2 (P <0 0 5 )。Western印迹检测结果表明 :2 0 μmol/Lgenistein处理SKOV3 细胞 12～ 72h后 ,c jun、c fos蛋白表达水平逐渐减弱。结论　genistein下调SKOV3 中EGFR介导的肿瘤信号转导通路中两个关键基因c erbB 2和c raf 1之mRNA及蛋白及其下游核转录因子c jun和c fos的mRNA及蛋白的表达水平 ,提示genistein干预EGFR介导的肿瘤信号转导系统中主要信号分子的表达可能是其抑制卵巢癌增殖的分子基础。Objective It was reported previously that genistein could inhibit proliferation of human ovarian carcinoma cell line SKOV 3, but mechanism was not clear There is a close relationship between EGFR mediated signal transduction pathway and the development of ovarian tumor This study aimed to investigate the effects of genistein on the EGFR mediated signal transduction pathway and to clarify its mechanisms of proliferation inhibition on human ovarian carcinoma cell line SKOV 3 and its xenograft in nude mice Methods The expression of c erbB 2 protein was determined using immunocytochemistry The expressions of c jun and c fos protein were determined using Western blotting The expression of c erbB 2, c raf 1, c jun and c fos mRNA were tested by reverse transcription polymerase chain reaction (RT PCR) Results The expression of c erbB 2, c raf 1 and its downstream gene c jun and c fos were decreased at mRNA level in the 20 μmol/L genistein group The expression of c erbB 2 protein were decreased, its average absorbency( A ) were decreased after treatment of SKOV 3 with 20 μmol/L genistein for 48 h, reached at 0 42 ±0 02( P <0 05) Western blotting demonstrated that the expressions of c jun and c fos protein were decreased gradually after being treated with 20 μmol/L genistein for 12-72 h Conclusions Genistein could down regulate the expression of two key genes,c erbB 2 and c raf 1 at mRNA and protein level in the EGFR mediated signal transduction pathway, and down regulate the expression of its downstream nuclear transcription factors c jun and c fos at mRNA and protein level It is suggested that interfering the expressions of some key signal molecules in EGFR mediated signal transduction system by genistein may account for its molecular foundation of proliferation inhibition in ovarian careinoma

关 键 词：c-jun 肿瘤 介导 SKOV3 裸鼠移植瘤 人卵巢癌细胞系 表达水平 印迹 RNA 信号转导通路 

分 类 号：R737.31[医药卫生—肿瘤] R735[医药卫生—临床医学]