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The carboxyl tail of Cx43 augments p38 mediated cell migration in a gap junction-independent manner

机译:Cx43的羧基尾以间隙连接独立的方式增强p38介导的细胞迁移

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摘要

The expression of connexin 43 (Cx43) has been shown to correlate with an enhanced migration of several cell types such as glioma or neural crest cells, but the mechanism remains unclear. We studied whether Cx43 also affects migration in non-neural cells and whether or not this is related to gap junction formation. Therefore, we analysed the migratory activity of HeLa cells under conditions of controlled connexin (Cx) expression. The expression of Cx43 enhanced their migration significantly as compared to Cx deficient wild-type cells. Expression of only the carboxyl tail of Cx43 (Cx43CT, AA 257-382) without channel forming capacity enhanced migration similarly as the full length protein. In contrast, the expression of the N-terminal part of Cx43 (Cx43NT, AA 1-257), which partially retained the gap junction channel function of Cx43, did not increase migration. The enhanced cell migration of HeLa cells expressing either full length Cx43 or the Cx43CT was associated with an increased activation of the p38 MAP kinase. The additional incubation with a specific inhibitor of p38 activation diminished the migration of HeLa-Cx43 cells to levels of control transfected cells.As a proof of concept, we studied whether Cx43 also modulates the migration of endothelial progenitor cells (EPC) which play an important role in angiogenesis. In these cells, which expressed Cx43 as the only connexin, the downregulation of Cx43 by siRNA resulted in a significantly decreased migration.These results demonstrate that expression of Cx43 augments migration via modulation of p38 MAP kinase activity. The carboxyl tail of Cx43 plays an essential role in this signalling pathway which is independent of gap junction function
机译:连接蛋白43(Cx43)的表达已显示与多种细胞类型(例如神经胶质瘤或神经correlat细胞)的迁移增强相关,但机制尚不清楚。我们研究了Cx43是否也影响非神经细胞的迁移,以及这是否与间隙连接的形成有关。因此,我们分析了受控连接蛋白(Cx)表达条件下HeLa细胞的迁移活性。与缺乏Cx的野生型细胞相比,Cx43的表达显着增强了它们的迁移。仅表达Cx43的羧基尾(Cx43CT,AA 257-382)而没有通道形成能力,与全长蛋白类似,其迁移增强。相反,部分保留了Cx43的间隙连接通道功能的Cx43(Cx43NT,AA 1-257)N端部分的表达并未增加迁移。表达全长Cx43或Cx43CT的HeLa细胞的细胞迁移增强与p38 MAP激酶激活增加有关。与特定的p38激活抑制剂共同孵育可减少HeLa-Cx43细胞向对照转染细胞水平的迁移。作为概念的证明,我们研究了Cx43是否还调节了起重要作用的内皮祖细胞(EPC)的迁移。在血管生成中的作用。在这些表达Cx43为唯一连接蛋白的细胞中,siRNA对Cx43的下调导致迁移明显减少。这些结果表明Cx43的表达通过调节p38 MAP激酶活性来增强迁移。 Cx43的羧基尾在该信号传导途径中起着至关重要的作用,而与间隙连接功能无关

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