Broberg A, Heino J
MediCity Research Laboratory and Department of Medical Biochemistry, University of Turku, Finland.
Exp Cell Res. 1996 Oct 10;228(1):29-35. doi: 10.1006/excr.1996.0295.
A cell culture inside a three-dimensional gel of fibrillar collagen is an experimental model used to study the response of cells to the extracellular matrix. Many cell types induce the contraction of gel and simultaneously decrease their production of type I collagen, whereas the expression of interstitial collagenase (matrix metalloproteinase-1; MMP-1) is enhanced. We have previously shown that in osteogenic cells the collagen receptor alpha2beta1 integrin is a positive regulator of MMP-1 and that the number of alpha2beta1 integrins on the cell surface also regulates the magnitude of contraction. However, the downregulation of collagen mRNA levels is not initiated by alpha2beta1 integrin. Here, we have studied in human KHOS-240 and MG-63 osteosarcoma cells and in human skin fibroblasts the effects of tyrosine kinase inhibitors on collagen gel contraction and on the regulation of MMP-1 and collagen alpha1(I) genes by extracellular collagen. The induction of MMP-1 could be inhibited by all tyrosine kinase inhibitors tested with the exception of genistein. None of them could prevent the downregulation of collagen expression. Thus, the collagen-induced alterations in the expression of MMP-1 and collagen alpha1(I) seem to be dependent on distinct signal transduction pathways. Many of the inhibitors, including genistein, could prevent the contraction of collagen gels. The effect was not related to their ability to inhibit cell growth, because an inhibitor specific for DNA synthesis and cell division did not have the same effect. Thus, we suggest that the process of collagen gel contraction requires protein-tyrosine phosphorylation and that the ability of cells to contract collagen gels is not related to the induction of MMP-1 or to the level of collagen alpha1(I) expression. Finally, we propose that the tyrosine kinase inhibitors might be considered as candidate molecules in the treatment of pathological scar contraction.
在纤维状胶原蛋白三维凝胶内的细胞培养是一种用于研究细胞对细胞外基质反应的实验模型。许多细胞类型会诱导凝胶收缩,同时减少其I型胶原蛋白的产生,而间质胶原酶(基质金属蛋白酶 -1;MMP -1)的表达则增强。我们之前已经表明,在成骨细胞中,胶原蛋白受体α2β1整合素是MMP -1的正向调节因子,并且细胞表面α2β1整合素的数量也调节收缩的程度。然而,胶原蛋白mRNA水平的下调不是由α2β1整合素启动的。在这里,我们研究了酪氨酸激酶抑制剂对人KHOS -240和MG -63骨肉瘤细胞以及人皮肤成纤维细胞中胶原蛋白凝胶收缩以及细胞外胶原蛋白对MMP -1和胶原蛋白α1(I)基因调节的影响。除金雀异黄素外,所有测试的酪氨酸激酶抑制剂都能抑制MMP -1的诱导。它们都不能阻止胶原蛋白表达的下调。因此,胶原蛋白诱导的MMP -1和胶原蛋白α1(I)表达的改变似乎依赖于不同的信号转导途径。许多抑制剂,包括金雀异黄素,都能阻止胶原蛋白凝胶的收缩。这种作用与其抑制细胞生长的能力无关,因为一种对DNA合成和细胞分裂具有特异性的抑制剂没有相同的作用。因此,我们认为胶原蛋白凝胶收缩过程需要蛋白质 - 酪氨酸磷酸化,并且细胞收缩胶原蛋白凝胶的能力与MMP -1的诱导或胶原蛋白α1(I)表达水平无关。最后,我们提出酪氨酸激酶抑制剂可能被视为治疗病理性瘢痕收缩的候选分子。