CCN2 promotes keratinocyte adhesion and migration via integrin α5β1
CCN2 promotes keratinocyte adhesion and migration via integrin α5β1
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DOI：
10.1016/j.yexcr.2013.08.021
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发表时间：
2013-11-15
影响因子：
3.7
通讯作者：
Eriksson, Elof
中科院分区：
文献类型：
--
作者：
Kiwanuka, Elizabeth;Andersson, Lauren;Eriksson, Elof
背景：CCN 2，（a.k.a.结缔组织生长因子和CTGF）已经成为细胞迁移的调节剂。虽然CCN 2对于伤口愈合中纤维化过程的重要性已经得到了很好的研究，但CCN 2对角质形成细胞功能的影响还没有得到很好的理解。在这项研究中，我们调查背后的CCN 2驱动的角质形成细胞粘附和migration.Materials和方法的机制：粘附测定进行涂布威尔斯孔与10 μ g/ml纤连蛋白（FN）或磷酸盐缓冲液（PBS）。在存在或不存在200 ng/ml CCN 2、5 mmol/l乙二胺四乙酸、10 mmol/l阳离子、500 μ l精氨酸-甘氨酸-天冬氨酸（RGD）、500 μ M精氨酸-甘氨酸-谷氨酸-丝氨酸（RGES）和10 μ g/ml抗整联蛋白封闭抗体的情况下接种角质形成细胞。使用改良的Boyden室测定法进行迁移研究。定量PCR检测CCN 2对整合素亚基mRNA表达的影响。为了阻断细胞内途径，在加入CCN 2之前，用20 μ M PD 98059（MEN-1抑制剂）或20 μ M PF 573228（FAN抑制剂）预处理角质形成细胞60分钟。Western blot检测CCN 2、p-ERK 1/2和ERK 1/2的表达。结果：CCN 2通过整合素α 5 β 1促进角质形成细胞与纤维连接蛋白的粘附。抗整合素α 5 β 1抗体的添加减少了CCN 2介导的角质形成细胞迁移。此外，CCN 2调节整合素亚基α 5和β 1的mRNA和蛋白质表达CCN 2激活FAK-MAPK信号通路，和预处理与MEK 1特异性抑制剂PD 98059显着减少CCN 2诱导的角质形成细胞migration.Conclusions：我们的研究结果表明，CCN 2增强角质形成细胞的粘附和迁移通过整合素α 5 β 1和激活FAK-MAPK信号级联。(C)2013 Elsevier Inc. All rights reserved.
Background: CCN2, (a.k.a. connective tissue growth factor and CTGF) has emerged as a regulator of cell migration. While the importance of CCN2 for the fibrotic process in wound healing has been well studied, the effect of CCN2 on keratinocyte function is not well understood. In this study, we investigated the mechanism behind CCN2-driven keratinocyte adhesion and migration.Materials and methods: Adhesion assays were performed by coating wells with 10 mu g/ml fibronectin (FN) or phosphate-buffered saline (PBS). Keratinocytes were seeded in the presence or absence of 200 ng/ml CCN2, 5 mmol/l ethylenediaminetetraacetic acid, 10 mmol/l cations, 500 mu l arginine-glycine-aspartic acid (RGD), 500 mu M arginine-glycine-glutamate-serine (RGES), and 10 mu g/ml anti-integrin blocking antibodies. Migration studies were performed using a modified Boyden chamber assay. Quantitative PCR was used to study the effect of CCN2 on integrin subunit mRNA expression. To block intracellular pathways, keratinocytes were pretreated with 20 mu M PD98059 (MEN-1 inhibitor) or 20 mu M PF573228 (FAN inhibitor) for 60 min prior the addition of CCN2. Western blot was used to measure CCN2, p-ERK1/2, and ERK1/2.Results: CCN2 enhanced keratinocyte adhesion to fibronectin via integrin alpha 5 beta 1. The addition of anti-integrin alpha 5 beta 1 antibodies reduced CCN2-mediated keratinocyte migration. In addition, CCN2 regulated mRNA and protein expression of integrin subunits alpha 5 and beta 1 CCN2 activated the FAK-MAPK signaling pathway, and pretreatment with MEK1-specific inhibitor PD98059 markedly reduced CCN2-induced keratinocyte migration.Conclusions: Our results demonstrate that CCN2 enhances keratinocyte adhesion and migration through integrin alpha 5 beta 1 and activation of the FAK-MAPK signaling cascade. (C) 2013 Elsevier Inc. All rights reserved.