Thrombin-induced CCN2 expression in human lung fibroblasts requires the c-Src/JAK2/STAT3 pathway
Thrombin-induced CCN2 expression in human lung fibroblasts requires the c-Src/JAK2/STAT3 pathway
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DOI：
10.1189/jlb.0911449
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发表时间：
2013-01-01
影响因子：
5.5
通讯作者：
Lin, Chien-Huang
中科院分区：
文献类型：
--
作者：
Bai, Kua-Jen;Chen, Bing-Chang;Lin, Chien-Huang
凝血酶是一种多功能丝氨酸蛋白酶，是诱导CCN2表达的重要纤维化介质。我们之前在人肺成纤维细胞中发现凝血酶通过ask1依赖的JNK/AP-1途径诱导CCN2表达。在这项研究中，我们进一步研究了c-Src、JAK2和STAT3在凝血素诱导的CCN2表达中的作用。凝血酶诱导的CCN2表达和CCN2- luc活性被JAK抑制剂（AG490）、JAK2DN、STAT3DN和STAT诱饵ODN减弱。此外，转染CCN2-mtSTAT-Luc构建体的细胞可以抑制凝血素诱导的CCN2-Luc活性。用凝血酶处理细胞导致JAK2 Tyr1007/1008位点磷酸化和STAT3 Tyr705位点磷酸化，并以时间依赖性的方式发生。凝血酶诱导的STAT3磷酸化被AG490和JAK2DN抑制。通过dna结合亲和力下拉试验分析凝血酶诱导的STAT3与CCN2启动子的结合。此外，凝血素诱导的CCN2表达和CCN2- luc活性被c-SrcDN和PP2（一种Src抑制剂）抑制。用c-SrcDN转染细胞也能抑制凝血酶诱导的JAK2和STAT3磷酸化。综上所述，这些结果表明凝血酶可能激活c-Src诱导JAK2活化，进而引起STAT3活化，最终诱导人肺成纤维细胞中CCN2的表达。j . Leukoc。生物学报。93:101-112；2013.
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. J. Leukoc. Biol. 93: 101-112; 2013.