Olsen D R, Peltonen J, Jaakkola S, Chu M L, Uitto J
Department of Dermatology, Jefferson Medical College, Philadelphia, PA 19107.
J Clin Invest. 1989 Mar;83(3):791-5. doi: 10.1172/JCI113959.
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.
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主要的胶原基因产物。在本研究中,通过分子杂交分析了培养的人皮肤成纤维细胞中10种不同基因的表达情况,这些基因编码I - VI型胶原的亚基多肽。Northern印迹分析表明存在I型、III型、IV型、V型和VI型胶原的特异性mRNA转录本,但不存在II型胶原的转录本。通过狭缝印迹杂交对这些mRNA丰度进行定量分析显示,I型、III型和VI型前胶原是培养的皮肤成纤维细胞主要的胶原基因产物。IV型和V型胶原的mRNA仅占总胶原mRNA转录本的一小部分。原位杂交进一步分析表明,大多数培养细胞共表达I型、III型和VI型前胶原原α链的基因。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,人皮肤成纤维细胞培养可用于研究至少9种遗传上不同的前胶原基因的转录调控。数据还进一步表明,除了I型和III型胶原外,VI型胶原可能是人皮肤的主要胶原成分。