IDENTIFICATION OF PROTEINS DIFFERENTIALLY EXPRESSED IN QUIESCENT AND PROLIFERATIVELY SENESCENT FIBROBLAST-CULTURES
IDENTIFICATION OF PROTEINS DIFFERENTIALLY EXPRESSED IN QUIESCENT AND PROLIFERATIVELY SENESCENT FIBROBLAST-CULTURES
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DOI：
10.1006/excr.1995.1304
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发表时间：
1995-09-01
影响因子：
3.7
通讯作者：
CRISTOFALO, VJ
中科院分区：
文献类型：
--
作者：
DIPAOLO, BR;PIGNOLO, RJ;CRISTOFALO, VJ
我们研究了静止（未生长）早期与晚期群体倍增水平（PDL） WI-38人二倍体成纤维细胞中蛋白质的差异表达。在分泌蛋白和总细胞蛋白水平上，基因表达的年龄相关变化通过高分辨率、二维凝胶电泳、n端测序或Western blot分析进行鉴定。分泌蛋白的比较揭示了衰老特异性缺失所有形式的EPC-1，一种与G(0)状态的早期PDL细胞相关的蛋白质。这一比较还揭示了衰老细胞中α I （III型）前胶原蛋白的加工（裂解）形式的缺失，表明细胞外基质的独特重塑。早期传代和增殖衰老成纤维细胞之间的细胞总蛋白比较发现，衰老细胞中泛素羧基末端水解酶的形式发生了改变，这一结果可以解释这些细胞中泛素化蛋白的增加。这些发现进一步阐明了体外衰老成纤维细胞的表型，并支持衰老细胞处于与G(0)早期PDL细胞根本不同的状态这一概念。(C) 1995学术出版社，Inc。
We have examined the differential expression of proteins in quiescent (nongrowing) early versus late population doubling level (PDL) WI-38 human diploid fibroblasts. Age-associated changes in gene expression at the level of both secreted and total cellular protein were identified using high-resolution, two-dimensional gel electrophoresis followed by N-terminal sequencing or Western blot analysis. A comparison of secreted proteins revealed the senescence-specific absence of all forms of EPC-1, a protein associated with early PDL cells in the G(0) state. This comparison also revealed the absence of a processed (cleaved) form of alpha I (type III) procollagen in senescent cells, suggesting a distinctive remodeling of the extracellular matrix. A comparison of total cellular protein between early passage and proliferatively aged fibroblasts identified an altered form of the enzyme ubiquitin carboxyl-terminal hydrolase in senescent cells, a result that could explain the increase in ubiquitinated proteins in these cells. These findings further elucidate the phenotype of fibroblasts aged in vitro and support the concept that senescent cells are arrested in a state fundamentally different from that of G(0) early PDL cells. (C) 1995 Academic Press, Inc.