干扰素γ抑制转化生长因子β，诱导的A549细胞上皮-间充质转化及对ERK1／2和STAT3的作用  

Interferon-γ inhibited epithelial-myofibroblast transition in A549 cells with upregulation of ERK1/2-STAT3 signalingsystem

作　　者：孙志宏[1] 许文兵[1] 朱元珏[1] 张宏 顾培 

机构地区：[1]北京协和医学院中国医学科学院北京协和医院呼吸科,100730 [2]北京中国医学科学院基础所细胞中心,100730

出　　处：《国际呼吸杂志》2012年第2期81-85,共5页International Journal of Respiration

摘　　要：目的探讨干扰素γ（IFN-γ）对转化生长因子β1（TGF-β1）诱导的A549细胞上皮-间充质转化（EMT）过程的作用及对胞外调节激酶1／2（ERK1／2）和信号转导和转录激活因子3（STAT3）的作用。方法①体外培养A549细胞，分为TGF-β1，处理组（5μg／L）、IFN-γ处理组（200μg／L）和TGF-1β（5μg／L）＋IFN-γ（10、100、200μg／L）联合处理组。处理时间48h，免疫荧光及Western blot方法检测各组细胞E-钙黏素（E-eadherin）、α-平滑肌肌动蛋白（α-SMA）、波形蛋白（vimentin）和纤维连接蛋白（fibronectin）的蛋白表达。②处理不同时间（5、30和60min），Western blot方法检测各组细胞STAT3、p—STAT3和ERK1／2、p-ERK1／2的蛋白表达。结果①与TGF-t31单独处理组相比，IFN-7（200μg／L）＋TGF-1β联合处理组vimentin和fibronectin的蛋白表达减低（P〈0．05），而α-SMA和E—cadherin蛋白表达无显著变化。②作用后5、30和60min，IFN-γ（200μg／L）＋TGF-1β联合处理组STAT3磷酸化水平显著升高（P〈0．01）。作用后5、60min，IFN-γ（200μg／L）＋TGF-1β联合处理组ERK1／2磷酸化水平升高（P〈0．05）。结论IFN-γ可在体外抑制TGF-1β。诱导的A549细胞发生EMT，其机制可能与上调ERK1／2和STAT3信号通路有关。Objective To examine the effects of interferon-γ （IFN-γ） on. epithelial-myofibroblast transition （EMT） of A549 cells induced by transforming growth faetor-β1 （TGF-β1） and its relation with extraeellular regulating kinase 1/2 （ERK1/2）-signal transducer and activator of transcription 3 （STAT3） signaling system. Methods Cultured A549 cells were divided into four groups：negative control, treated with TGF-β1 （5 μg/L） as positive control,treated with IFN-γ （200 μg/L）,co-treated with TGF-β1 （5 μg/L） and IFN-γ （10,100,200 μg/L） for 48 hours. The protein expressions of E-cadherin, α-smooth muscle actin （α-SMA）,vimentin and fibronectin were assessed by indirect immunofluorescence or Western blot, respectively. In the other experiment,A549 cells were co-treated with IFN-γ （200 μg/L） and TGF-β1 for different time,respectively. The protein expressions of STAT3, p-STAT3, ERK1/2 and p-ERK1/2 were detected by Western blot. Results IFN-γ （200μg/L） reversed the promotion of vimentin and fibronectin protein expressions induced by TGF-β1 significantly （ P 〈0.05）, whereas both α-SMA and E-cadherin protein expressions changed a little. The ratio of p-STAT3 and STAT3 was significantly increased in group co-treated with TGF-α and IFN-γ （200 μg/L） at 5,30 and 60 min as compared with positive controls （ P 〈0.01）. At the same time, the ratio of p-ERK1/2 and ERK1/2 was significantly increased at 5 and 60 min （ P 〈0.05）. Conclusions IFN-γ may inhibit EMT in A549 cells induced by TGF-β1. This effect may involve upreguiation of ERK1/2-STAT3 signaling system.

关 键 词：干扰素Γ 上皮-间充质转化 胞外调节激酶1/2 信号转导和转录激活因子3 

分 类 号：R363[医药卫生—病理学]