Smad7基因对胞外信号调节激酶磷酸化水平的调控  

Smad7 Regulates the Phosphorylation of Extracellular Signal-Regulated Kinase

作　　者：霍艳英[1] 李刚[1] 王莹[2] 张开泰[1] 何欣蓉[2] 刘敏丽[2] 项晓琼[1] 胡迎春[1] 吴德昌[1] 

机构地区：[1]军事医学科学院放射医学研究所,北京100850 [2]重庆医科大学病理学教研室,重庆400016

出　　处：《生物技术通讯》2005年第3期239-241,共3页Letters in Biotechnology

基　　金：国家自然科学基金面上项目(30200329)

摘　　要：研究在BEP2D细胞中,作为Smads蛋白家族的抑制分子,Smad7对胞外信号调节的蛋白激酶(ERK1/2或p44/42)磷酸化水平的调控。将Smad7真核表达载体或人工合成的Smad7-siRNA转染BEP2D细胞,TGF-β刺激,通过Western印迹检测Smad7对p44/42蛋白磷酸化的影响。结果在永生化BEP2D细胞中,TGF-β1刺激后5min开始,可以检测到磷酸化的p44/42;到60min达到高峰,之后逐渐降低。细胞转染Smad7,TGF-β作用60min后,p44/42磷酸化水平明显增高;而转染Smad7-SiRNA,TGF-β作用60min后,p44/42磷酸化水平显著降低。p44/42蛋白水平基本上不受TGF-β1刺激及Smad7表达水平的影响。以上结果说明,在BEP2D细胞中,Smad7可参与TGF-β对ERK/MAPK通路的活化作用。To analyze the regulation effect of Smad7 gene on phosphorylation of extracellular signal-regulated protein kinase(ERK) in the immortalized human broncbial epithelial cells BEP2D. Full-length Smad7 cDNA construct pCISmad7.neo or Smad7-SiRNA was tranfected into a mammalian cell line BEP2D, which was then stimulated by TGF-β1. Western blots were performed to investigate the regulation effect of Smad7 on ERK signal pathway. In BEP2D cells, activations of p42 and p44 were detected 5 min after TGF-β1 treatment, with peak activation occurring at 60 min, then decreased later. Transient transfection of pCISmad7.neo led to increased activation of p44/42. Transient transfection of Smad7-SiRNA led to decreased activition of p44/42. The protein level of p42 and p44 were not significantly affected by TGF-β1 treatment or the expression level of Smad7. In BEP2D cells, Smad7 participates in the activation of ERK/MAPK signal pathway mediated by TGF-β1.

关 键 词：信号调节 调控 WESTERN印迹 Smad7 SMADS蛋白 基因 ERK1/2 真核表达载体 TGF-β1 蛋白磷酸化 D细胞 蛋白激酶 人工合成 细胞转染 蛋白水平 活化作用 MAPK β作用 永生化 酸化水 检测 

分 类 号：Q786[生物学—分子生物学] TP212[自动化与计算机技术—检测技术与自动化装置]