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培养的黑色素瘤细胞的表型分析。M5人黑色素瘤细胞系中细胞角蛋白型中间丝的表达。

Phenotypic analysis of cultured melanoma cells. Expression of cytokeratin-type intermediate filaments by the M5 human melanoma cell line.

作者信息

Trejdosiewicz L K, Southgate J, Kemshead J T, Hodges G M

出版信息

Exp Cell Res. 1986 Jun;164(2):388-98. doi: 10.1016/0014-4827(86)90037-6.

Abstract

Expression of intermediate filament (IF) isotypes was studied in six human and two murine melanoma cell lines. With one exception, these lines expressed IFs only of the vimentin type; neurofilament peptides, desmin and GFAP were not detected. However, the M5 human melanoma line also expressed extensive cytokeratin tonofilament arrays, as visualized by immunofluorescence with a panel of eleven monoclonal antibodies and hetero-antisera to cytokeratins; only the keratin 19-specific antibody BA16 did not react. By 2 D gel electrophoresis, five major keratin peptides were detected (keratins 7, 8, 13, 17 and 18), and an additional 57 kD peptide was detected on immunoblots with several antikeratin antibodies. Also observed in M5 cells was focal collapse of tonofilament arrays in mitotic cells. All the melanoma lines tested were positive for S100; M5 and two other cell lines were also positive for the 220-240 kD neuroectoderm-associated cell-surface differentiation antigen defined by monoclonal antibody UJ 127:11. In all the melanoma cell lines, secretion of extracellular matrix proteins (fibronectin, laminin and collagen type IV) was sparse or absent, and all were negative for the epithelial cell markers HMG-1 and HMG-2. Co-expression of keratin and vimentin by a melanoma cell line is discussed in the light of recent controversy concerning expression of cytokeratins by other neoplasms of putative neuroectodermal origins.

摘要

在六种人类和两种小鼠黑色素瘤细胞系中研究了中间丝(IF)同种型的表达。除一个例外,这些细胞系仅表达波形蛋白类型的中间丝;未检测到神经丝肽、结蛋白和胶质纤维酸性蛋白(GFAP)。然而,M5人黑色素瘤细胞系也表达广泛的细胞角蛋白张力丝阵列,这通过用一组11种单克隆抗体和针对细胞角蛋白的异种抗血清进行免疫荧光观察得以证实;只有角蛋白19特异性抗体BA16没有反应。通过二维凝胶电泳,检测到五种主要的角蛋白肽(角蛋白7、8、13、17和18),并且在用几种抗角蛋白抗体进行的免疫印迹上检测到一种额外的57kD肽。在M5细胞中还观察到有丝分裂细胞中张力丝阵列的局灶性塌陷。所有测试的黑色素瘤细胞系S100均呈阳性;M5和另外两种细胞系对于单克隆抗体UJ 127:11所定义的220 - 240kD神经外胚层相关细胞表面分化抗原也呈阳性。在所有黑色素瘤细胞系中,细胞外基质蛋白(纤连蛋白、层粘连蛋白和IV型胶原)的分泌稀少或不存在,并且所有细胞系上皮细胞标志物HMG - 1和HMG - 2均为阴性。鉴于最近关于假定神经外胚层起源肿瘤细胞角蛋白表达的争议,讨论了黑色素瘤细胞系中角蛋白和波形蛋白的共表达情况。

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