Collagen gene expression by cultured human skin fibroblasts. Abundant steady-state levels of type VI procollagen messenger RNAs.
Collagen gene expression by cultured human skin fibroblasts. Abundant steady-state levels of type VI procollagen messenger RNAs.
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DOI：
10.1172/jci113959
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发表时间：
1989-03
期刊：
影响因子：
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通讯作者：
David R. Olsen;Juha Peltonen;S. Jaakkola;Mon-Li Chu;Jouni Uitto
中科院分区：
文献类型：
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作者：
David R. Olsen;Juha Peltonen;S. Jaakkola;Mon-Li Chu;Jouni Uitto
以往的研究表明，I型和III型前胶原是培养的人皮肤成纤维细胞的主要胶原基因产物。本研究用分子杂交的方法分析了人皮肤成纤维细胞中编码I-VI型胶原亚基多肽的10个不同基因的表达。Northern Transfer分析表明，I型、III型、IV型、V型和VI型胶原存在特异的mRNA转录本，而II型胶原没有。缝隙印迹杂交结果表明，I、III、VI型前胶原是皮肤成纤维细胞的主要胶原基因产物。IV型和V型胶原蛋白的mRNAs仅占总胶原蛋白mRNA转录本的一小部分。进一步的原位杂交分析表明，大多数培养细胞共表达I型、III型和VI型前胶原α链基因。进一步的原位杂交分析揭示了VI型胶原基因在正常人皮肤中的表达。这些数据表明，人类皮肤成纤维细胞培养可以用来研究至少九个遗传上不同的前胶原基因的转录调控。这些数据进一步表明，除了I型和III型胶原外，VI型胶原可能是人类皮肤的主要胶原成分。
Previous studies have suggested that procollagen types I and III are the major collagenous gene products of cultured human skin fibroblasts. In this study the expression of 10 different genes, encoding the subunit polypeptides for collagen types I-VI, by human skin fibroblasts in culture was analyzed by molecular hybridizations. Northern transfer analysis demonstrated the presence of specific mRNA transcripts for collagen types I, III, IV, V, and VI, but not for type II collagen. Quantitation of the abundance of these mRNAs by slot blot hybridizations revealed that type I, III, and VI procollagens were the major collagenous gene products of skin fibroblasts in culture. The mRNAs for type IV and V collagens represented only a small percentage of the total collagenous mRNA transcripts. Further analysis by in situ hybridization demonstrated that the majority of the cultured cells coexpressed the genes for type I, III, and VI procollagen pro-alpha chains. Further in situ hybridization analyses revealed the expression of type VI collagen genes in normal human skin. These data demonstrate that human skin fibroblast cultures can be used to study the transcriptional regulation of at least nine genetically distinct procollagen genes. The data further suggest that type VI collagen, in addition to types I and III, may be a major collagenous component of human skin.