CAJ | 학술논문

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(5 μg/L)、IFN-γ处理组(200 μg/L)和TGF-β1(5 μg/L)+ IFN-γ(10、100、200 μg/L)联合处理组.处理时间48 h,免疫荧光及Western blot方法检测各组细胞E-钙黏素( E-cadherin)、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60 min),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-β1单独处理组相比,IFN-γ(200 μg/L)+TGF-β1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60 min,IFN-γ(200 μg/L)+ TGF-β1联合处理组STAT3磷酸化水平显著升高(P＜0.01).作用后5、60 min,IFN-γ(200 μg/L) +TGF-β1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-β1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.
목적 탐토간우소γ(IFN-γ)대전화생장인자β1(TGF-β1)유도적A549세포상피-간충질전화(EMT)과정적작용급대포외조절격매1/2(ERK1/2)화신호전도화전록격활인자3(STAT3)적작용.방법 ①체외배양A549세포,분위TGF-β1처리조(5 μg/L)、IFN-γ처리조(200 μg/L)화TGF-β1(5 μg/L)+ IFN-γ(10、100、200 μg/L)연합처리조.처리시간48 h,면역형광급Western blot방법검측각조세포E-개점소( E-cadherin)、α-평활기기동단백(α-SMA)、파형단백(vimentin)화섬유련접단백(fibronectin)적단백표체.②처리불동시간(5、30화60 min),Western blot방법검측각조세포STAT3、p-STAT3화ERK1/2、p-ERK1/2적단백표체.결과 ①여TGF-β1단독처리조상비,IFN-γ(200 μg/L)+TGF-β1연합처리조vimentin화fibronectin적단백표체감저(P＜0.05),이α-SMA화E-cadherin단백표체무현저변화.②작용후5、30화60 min,IFN-γ(200 μg/L)+ TGF-β1연합처리조STAT3린산화수평현저승고(P＜0.01).작용후5、60 min,IFN-γ(200 μg/L) +TGF-β1연합처리조ERK1/2린산화수평승고(P＜0.05).결론 IFN-γ가재체외억제TGF-β1유도적A549세포발생EMT,기궤제가능여상조ERK1/2화STAT3신호통로유관.
Objective To examine the effects of interferon-γ (IFN-γ) on epithelial-myofibroblast transition (EMT) of A549 cells induced by transforming growth factor-β1 (TGF-β1) and its relation with extracellular 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 (200 μg/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-β1 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 upregulation of ERK1/2-STAT3 signaling system.