Mitogen-activated protein kinase activation mediates PDGF-directed migration of RPE cells
Mitogen-activated protein kinase activation mediates PDGF-directed migration of RPE cells
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DOI：
10.1006/excr.1997.3873
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发表时间：
1998-02-25
影响因子：
3.7
通讯作者：
Law, RE
中科院分区：
文献类型：
--
作者：
Hinton, DR;He, SK;Law, RE
生长因子导向的迁移是伤口愈合反应的关键组成部分，尽管对涉及的信号通路知之甚少。为研究丝裂原活化蛋白激酶(MAPK)通路对血小板衍生生长因子(PDGF)和纤维连接蛋白诱导的人视网膜色素上皮(RPE)细胞迁移的影响，采用Transwell细胞培养箱，观察PDGF-BB(10-50 ng/ml)和纤维连接蛋白(FN)对有或无MAPK通路抑制剂PD98059(10-30mU M)对RPE细胞迁移成分的影响。免疫沉淀法和免疫组织化学方法检测PDGF-BB对血清饥饿细胞MAPK的激活作用。PDGF-BB(10 ng/mI)刺激RPE的MAPK活性(10min)及其核定位(1h)。PD98059抑制PDGF-BB或血清激活的MAPK，PDGF-BB刺激RPE的趋化、趋化和趋化；纤维连接蛋白底物可增加趋化作用并协同增强趋化作用，PD98059可有效抑制纤维连接蛋白诱导的趋化作用和PDGF-BB诱导的趋化作用，但只有较高浓度的PDGF-BB(50 ng/ml)才能抑制趋化作用。这些结果表明，MAPK在RPE体外迁移的多种成分中起关键作用，并提示靶向MAPK在体内抑制RPE迁移的可能性。(C)1998年学术出版社。
Growth factor-directed migration is a critical component of the wound healing response although little is known about the signaling pathways involved. We examined the effect of inhibiting the mitogen-activated protein kinase (MAPK) pathway on platelet-derived growth factor (PDGF) and fibronectin-induced cell migration of human retinal pigment epithelial (RPE) cells, Using transwell cell-culture chambers, the effect of PDGF-BB (10-50 ng/ml) and fibronectin on components of migration was measured with or without the MAPK pathway inhibitor PD98059 (10-30 mu M). MAPK activation of serum-starved cells by PDGF-BB was demonstrated by an immunoprecipitation/kinase assay and by immunohistochemistry using antibody specific for phosphorylated MAPK. PDGF-BB (10 ng/mi) stimulated MAPK activity in RPE (10 min) and its nuclear localization (1 h). PD98059 inhibited the activation of MAPK by PDGF-BB or serum, PDGF-BB stimulated RPE chemokinesis, chemotaxis, and haptotaxis; chemokinesis was additively increased and chemotaxis synergistically increased by the presence of a fibronectin substratum, PD98059 potently inhibited fibronectin-induced haptotaxis and PDGF-BB-induced chemotaxis but inhibited chemokinesis only at higher PDGF-BB (50 ng/ml) concentrations in the presence of fibronectin substratum. These results demonstrate that MAPK is critically involved in multiple components of RPE migration in vitro and suggest the potential of targeting MAPK to inhibit RPE migration in vivo. (C) 1998 Academic Press.