PDF(Pubmed)
Abstract：
Vimentin has been reported to play diverse roles in cell processes such as spreading, migration, cell-matrix adhesion, and fibrotic transformation. Here, we assess how vimentin impacts cell spreading, morphology, and myofibroblast transformation of human corneal fibroblasts. Overall, although knockout (KO) of vimentin did not dramatically impact corneal fibroblast spreading and mechanical activity (traction force), cell elongation in response to PDGF was reduced in vimentin KO cells as compared to controls. Blocking vimentin polymerization using Withaferin had even more pronounced effects on cell spreading and also inhibited cell-induced matrix contraction. Furthermore, although absence of vimentin did not completely block TGFβ-induced myofibroblast transformation, the degree of transformation and amount of αSMA protein expression was reduced. Proteomics showed that vimentin KO cells cultured in TGFβ had a similar pattern of protein expression as controls. One exception included periostin, an ECM protein associated with wound healing and fibrosis in other cell types, which was highly expressed only in Vim KO cells. We also demonstrate for the first time that LRRC15, a protein previously associated with myofibroblast transformation of cancer-associated fibroblasts, is also expressed by corneal myofibroblasts. Interestingly, proteins associated with LRRC15 in other cell types, such as collagen, fibronectin, β1 integrin and α11 integrin, were also upregulated. Overall, our data show that vimentin impacts both corneal fibroblast spreading and myofibroblast transformation. We also identified novel proteins that may regulate corneal myofibroblast transformation in the presence and/or absence of vimentin.
摘要：
据报道，波形蛋白在细胞过程中起着不同的作用，例如传播，迁移,细胞-基质粘附，和纤维化转化。这里,我们评估波形蛋白如何影响细胞扩散，形态学,和人角膜成纤维细胞的肌成纤维细胞转化。总的来说,尽管波形蛋白的敲除（KO）并没有显着影响角膜成纤维细胞的扩散和机械活动（牵引力），与对照相比,波形蛋白KO细胞中响应于PDGF的细胞伸长降低。使用Withaferin阻断波形蛋白聚合对细胞扩散具有更明显的影响，并且还抑制了细胞诱导的基质收缩。此外，尽管缺乏波形蛋白并不能完全阻断TGFβ诱导的肌成纤维细胞转化，转化程度和αSMA蛋白表达量降低。蛋白质组学显示在TGFβ中培养的波形蛋白KO细胞具有与对照相似的蛋白质表达模式。一个例外包括骨膜素，与其他细胞类型的伤口愈合和纤维化相关的ECM蛋白，仅在VimKO细胞中高度表达。我们还首次证明了LRRC15，一种先前与癌症相关成纤维细胞转化相关的蛋白，也由角膜肌成纤维细胞表达。有趣的是,其他细胞类型中与LRRC15相关的蛋白质，如胶原蛋白，纤连蛋白,β1整合素和α11整合素，也被上调了。总的来说,我们的数据显示波形蛋白影响角膜成纤维细胞扩散和肌成纤维细胞转化.我们还鉴定了在存在和/或不存在波形蛋白的情况下可能调节角膜肌成纤维细胞转化的新型蛋白质。
