Experimental coexpression of vimentin and keratin intermediate filaments in human melanoma cells augments motility.
Experimental coexpression of vimentin and keratin intermediate filaments in human melanoma cells augments motility.
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发表时间：
1996
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通讯作者：
Yi Wen Chu;E. Seftor;Lewis H. Romer;M. J. Hendrix
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作者：
Yi Wen Chu;E. Seftor;Lewis H. Romer;M. J. Hendrix
中间丝已被用作分化和病理学中的细胞类型特异性标志物;然而，最近的报告已经证明了波形蛋白（间充质标志物）和角蛋白（上皮标志物）在许多肿瘤中的共表达，包括与转移性疾病有关的黑色素瘤。为了检验黑色素瘤细胞共表达波形蛋白和角蛋白有助于更具迁移性和侵袭性表型的假设，我们用角蛋白8和18的cDNA共转染波形蛋白阳性的人黑色素瘤细胞系A375 P（低侵袭能力）。所得到的稳定的转染表达波形蛋白和角蛋白阳性的中间丝表现出两到三倍的增加，他们的基底膜基质和迁移，通过明胶在体外的入侵。这些发现得到了电影录像的进一步证实。在纤连蛋白上附着和铺展期间，含有波形蛋白和角蛋白8和18的转染子表现出对β 1整联蛋白和磷酸酪氨酸染色阳性的局灶性粘附的增加，沿着增强的膜皱褶和肌动蛋白应力纤维形成。从这些数据中，我们推测，波形蛋白和角蛋白的共表达导致细胞骨架相互作用增加，在涉及整合素细胞信号传导事件的细胞外基质内的焦点接触，这有助于更多的迁移行为。
Intermediate filaments have been used as cell-type-specific markers in differentiation and pathology; however, recent reports have demonstrated the coexpression of vimentin (a mesenchymal marker) and keratins (epithelial markers) in numerous neoplasms, including melanoma, which has been linked to metastatic disease. To test the hypothesis that coexpression of vimentin and keratins by melanoma cells contributes to a more migratory and invasive phenotype, we co-transfected a vimentin-positive human melanoma cell line, A375P (of low invasive ability), with cDNAs for keratins 8 and 18. The resultant stable transfectants expressed vimentin- and keratin-positive intermediate filaments showed a two- to threefold increase in their invasion of basement membrane matrix and migration through gelatin in vitro. These findings were further corroborated by video cinematography. During attachment and spreading on fibronectin, the transfectants containing vimentin and keratins 8 and 18 demonstrated an increase in focal adhesions that stained positive for beta 1 integrin and phosphotyrosine, along with enhanced membrane ruffling and actin stress fiber formation. From these data, we postulate that coexpression of vimentin and keratins results in increased cytoskeletal interactions at focal contacts within extracellular matrices involving integrin cell signaling events, which contributes to a more migratory behavior.