Codistribution of pericellular matrix proteins in cultured fibroblasts and loss in transformation: fibronectin and procollagen.
Codistribution of pericellular matrix proteins in cultured fibroblasts and loss in transformation: fibronectin and procollagen.
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培养的成纤维细胞中细胞周基质蛋白的共分布和转化过程中的损失：纤连蛋白和前胶原。
DOI：
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发表时间：
1978
影响因子：
11.1
通讯作者：
R. Timpl
中科院分区：
文献类型：
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作者：
Antti Vaheri;Markku Kurkinen;V.;Ewert Linder;R. Timpl
纤连蛋白和不同类型的前胶原和胶原蛋白的抗体被用于免疫荧光染色，以定位这些蛋白质在细胞培养物中。正常人皮肤或肺成纤维细胞产生的纤维细胞周围基质中，纤连蛋白和前胶原（I型和III型）表现出广泛的共分布。纤维连接蛋白和前胶原由相同的细胞合成，通过双染色免疫荧光判断。胶原酶特异性消化细胞周前胶原，而不影响基质纤维连接蛋白的纤维状分布。用胰蛋白酶的简单处理去除了两种基质蛋白。人肿瘤细胞系HT-1080（纤维肉瘤）和RD（横纹肌肉瘤）产生很少或不产生基质纤连蛋白或前胶原。在细胞接触部位，猴病毒40转化的肺成纤维细胞（VA 13）产生少量与I型前胶原共分布的细胞周纤维基质纤连蛋白。在所有这些人肉瘤细胞系中可见细胞内纤维连接蛋白和前胶原。当鸡胚成纤维细胞感染的T类突变体（NY 68）劳斯肉瘤病毒的温度敏感的转化保持在非允许的温度（41度）的细胞有正常的表型和纤维状基质含有纤连蛋白和原胶原蛋白。在允许温度（35 ℃）下，细胞表现出转化的表型，基质丢失。不能产生细胞周围纤连蛋白/胶原基质可能是转化培养成纤维细胞的几种表型特征的原因。
Antibodies to fibronectin and to distinct types of procollagens and collagens were used in immunofluorescent staining to localize these proteins in cell cultures. Normal human skin or lung fibroblasts produced a fibrillar pericellular matrix in which fibronectin and procollagen (types I and III) showed extensive codistribution. Fibronectin and procollagen were synthesized by the same cells as judged by double-stain immunofluorescence. Pericellular procollagen was specifically digested with collagenase without an effect on the fibrillar distribution of matrix fibronectin. Brief treatment with trypsin removed both matrix proteins. The human tumor cell lines HT-1080 (fibrosarcoma) and RD (rhabdomyosarcoma) produced little or no matrix fibronectin or procollagen. At sites of cell contact, simian virus 40-transformed lung fibroblasts (VA13) produced small amounts of pericellular fibrillar matrix fibronectin that codistributed with procollagen type I. Intracellular fibronectin and procollagen were visualized in all of these human sarcoma cell lines. When chicken embryo fibroblasts infected with a T class mutant (NY68) of Rous sarcoma virus temperature-sensitive for transformation were maintained at the nonpermissive temperature (41 degrees ) the cells had normal phenotype and a fibrillar matrix containing fibronectin and procollagen was present. At the permissive temperature (35 degrees ), the cells showed transformed phenotype and the matrix was lost. The failure to produce a pericellular fibronectin/collagen matrix may account for several phenotypic characteristics of transformed cultured fibroblasts.