Thrombin-induced CCN2 expression in human lung fibroblasts requires the c-Src/JAK2/STAT3 pathway

Journal of Leukocyte Biology

Kua-Jen Bai, ⁴  Chung-Chi Yu³  Chih-Hsiung Wu–¹  Hon-Ping Ma–¹  Chuang-Ye Hong⁶  Hui-Chen Pai³  Ming-Jen Hsu³  Ming-Chih Yu⁴  Chien-Huang Lin,–²  Chih-Ming Weng³  Bing-Chang Chen ⁵  Min-Liang Kuo and⁷ 

[1] –Taipei Medical University“Shuang Ho Hospital, New Taipei City, Taiwan;Department of Pulmonary Medicine, Taipei Medical University“Wanfang Hospital, and Graduate Institute of Medical Sciences, and –Taipei Medical University“Shuang Ho Hospital, New Taipei City, Taiwan;Graduate Institute of Medical Sciences, and Department of Pharmacology, College of Medicine, Taipei Medical University, Taipei, Taiwan;Department of Pulmonary Medicine, Taipei Medical University“Wanfang Hospital, and  School of Respiratory Therapy, Department of Pharmacology, College of Medicine, Taipei Medical University, Taipei, Taiwan; School of Respiratory Therapy, Department of Pharmacology, College of Medicine, Taipei Medical University, Taipei, Taiwan;Department of Pulmonary Medicine, Taipei Medical University“Wanfang Hospital, and Department of Pharmacology, College of Medicine, Taipei Medical University, Taipei, Taiwan;Angiogenesis Research Center, Laboratory of Molecular and Cellular Toxicology, Institute of Toxicology, College of Medicine, National Taiwan University, Taipei, Taiwan Department of Pharmacology, College of Medicine, Taipei Medical University, Taipei, Taiwan;

关键词: inflammation;    transcription factors;    gene regulation;    signal transduction;    lung;   

DOI  :  10.1189/jlb.0911449

学科分类：生理学

来源: Federation of American Societies for Experimental Biology

【 摘 要 】

Thrombin is a multifunctional serine protease and an important fibrotic mediator that induces CCN2 expression. We previously showed that thrombin induces CCN2 expression via an ASK1-dependent JNK/AP-1 pathway in human lung fibroblasts. In this study, we further investigated the roles of c-Src, JAK2, and STAT3 in thrombin-induced CCN2 expression. Thrombin-induced CCN2 expression and CCN2-Luc activity were attenuated by a JAK inhibitor (AG490) and JAK2DN, STAT3DN, and the STAT decoy ODN. Moreover, transfection of cells with a CCN2-mtSTAT-Luc construct inhibited thrombin-induced CCN2-Luc activity. Treatment of cells with thrombin caused JAK2 phosphorylation at Tyr1007/1008 and STAT3 phosphorylation at Tyr705 in time-dependent manners. Thrombin-induced STAT3 phosphorylation was inhibited by AG490 and JAK2DN. Thrombin-induced STAT3 binding to the CCN2 promoter was analyzed by a DNA-binding affinity pull-down assay. In addition, thrombin-induced CCN2 expression and CCN2-Luc activity were inhibited by c-SrcDN and PP2 (an Src inhibitor). Transfection of cells with c-SrcDN also inhibited thrombin-induced JAK2 and STAT3 phosphorylation. Taken together, these results indicate that thrombin might activate c-Src to induce JAK2 activation, which in turn, causes STAT3 activation, and finally induces CCN2 expression in human lung fibroblasts.

【 授权许可】

Unknown