β-雌二醇经ERβ-ERK1/2雌激素胞膜信号传导途径促进肺癌A549细胞迁移和侵袭  

β-estradiol promotes invasion and migration of lung cancer A549 cells via ERβ-mediated ERK1/2 membrane-initiated steroid signaling

作　　者：董年 陈俊杰[1] 石林 陈成水[1] DONG Nian;CHEN Jun-jie;SHI Lin;CHEN Cheng-shui(Department of Respiratory and Critical Care Medicine,The First Affiliated Hospital,Wenzhou Medical University,Wenzhou 325000,China;Department of Respiratory,Zhongshan Hospital,Fudan University,Shanghai 200000,China.)

机构地区：[1]温州医科大学附属第一医院呼吸与危重症医学科,浙江温州325000 [2]复旦大学附属中山医院呼吸内科,上海200000

出　　处：《中国病理生理杂志》2018年第10期1742-1747,共6页Chinese Journal of Pathophysiology

基　　金：浙江省卫生厅项目(No.2015KYB246);温医一院院级科研孵化课题(No.FHY2015037)

摘　　要：目的:研究β-雌二醇(β-estradiol)促进肺癌A549细胞迁移和侵袭的调控机制。方法:体外培养乳腺癌MCF-7细胞和肺癌A549细胞,以MCF-7细胞作为阳性对照明确雌激素受体(ER)在A549细胞中的表达情况; real-time PCR检测ERβ亚型ERβ1、ERβ2和ERβ5在A549细胞中的表达情况,免疫荧光定位ERβ在细胞内的胞膜和胞核分布情况;予β-estradiol刺激A549细胞,Western blot检测胞内ERK1/2磷酸化水平改变,Transwell小室侵袭实验和细胞实时监测培养系统检测细胞迁移和侵袭能力的变化;利用ERK1/2抑制剂PD98059预先处理,Transwell小室侵袭实验和细胞实时监测培养系统检测β-estradiol刺激后细胞迁移和侵袭能力的变化。结果:ER在A549细胞中以表达ERβ为主,且表达的ERβ以ERβ2和ERβ5为主,免疫荧光显示细胞内ERβ以胞质分布为主。β-estradiol可以引起A549细胞ERK1/2磷酸化水平的增加,并促进细胞的迁移和侵袭,抑制ERK1/2信号传导可以逆转β-estradiol促进的细胞迁移和侵袭。结论:β-estradiol经ERβ活化ERK1/2雌激素胞膜信号传导,从而促进A549细胞的侵袭和转移。ERK1/2是ERβ雌激素胞膜信号传导中与肺癌侵袭和转移相关的重要信号节点。AIM:To investigate the regulatory mechanism ofβ-estradiol in the invasion and migration of lung cancer A549 cells.METHODS:Breast cancer MCF-7 cells and lung cancer A549 cells were cultured in vitro.The MCF-7 cells were used as the estrogen receptor(ER)positive expression cell model.Real-time PCR and immunofluorescence were employed to measure the expression level and the localization of ER in A549 cells.The phosphorylation of ERK1/2 uponβ-estradiol stimulation was quantified by Western blot.The invasion and migration abilities of A549 cells uponβ-estradiol stimulation with or without ERK1/2 inhibitor PD98059 were measured by Transwell and Cell-IQ assays.RESULTS:ERβwas the dominant ER subtype in the A549 cells and primarily comprised of ERβ2 and ERβ5.Immunofluorescence revealed that ERβexpression was mainly localized in the cytoplasm.β-estradiol induced phosphorylation of ERK1/2 and promoted the invasion and migration of the cells.Inhibition of ERK1/2 signaling reversedβ-estradiol-promoted invasion and migration of A549 cells.CONCLUSION:ERβ-mediated membrane-initiated steroid signaling is involved in the process ofβ-estradiol-promoted invasion and migration of A549 cells,through which ERK1/2 signaling plays a pivotal role.

关 键 词：肺癌 Β-雌二醇 雌激素受体 ERK1/2信号通路 细胞迁移 肿瘤侵袭 

分 类 号：R329.28[医药卫生—人体解剖和组织胚胎学] R734.2[医药卫生—基础医学]