Targeted disruption of the tissue inhibitor of metalloproteinases gene increases the invasive behavior of primitive mesenchymal cells derived from embryonic stem cells in vitro.
Targeted disruption of the tissue inhibitor of metalloproteinases gene increases the invasive behavior of primitive mesenchymal cells derived from embryonic stem cells in vitro.
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DOI：
10.1083/jcb.118.3.727
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发表时间：
1992-08
期刊：
影响因子：
--
通讯作者：
Werb Z
中科院分区：
文献类型：
--
作者：
Alexander CM;Werb Z
金属蛋白酶家族的蛋白水解酶可以降解细胞外基质并促进侵袭性迁移。这类酶被金属蛋白酶组织抑制剂（TIMP-1）特异性抑制。利用同源重组，我们已经破坏了多能胚胎干细胞中编码TIMP-1的基因。由于TIMP-1基因是X连锁的，并且在胚胎干细胞中是半合子的，我们已经能够在培养中研究这种突变的影响。使用基底膜侵袭试验，我们发现，在低浓度的血清与维甲酸分化的突变细胞，比他们的正常细胞的同行更具侵袭性，这是专门逆转加入外源性TIMP-1蛋白。侵袭性细胞群体具有原始间充质细胞的早期群体的特征，包括波形蛋白的表达和开始分化后4-8 d的短暂侵袭期。因此，金属蛋白酶活性可以是细胞侵袭的速率限制。
The metalloproteinase family of proteolytic enzymes can degrade extracellular matrix and facilitate invasive migration. This class of enzymes is specifically inhibited by the tissue inhibitor of metalloproteinases (TIMP-1). Using homologous recombination, we have disrupted the gene encoding TIMP-1 in pluripotent embryonic stem cells. Because the TIMP-1 gene is X linked and is hemizygous in embryonic stem cells, we have been able to study the effect of this mutation in culture. Using a basement membrane invasion assay, we found that the mutant cells, differentiated in low concentrations of serum with retinoic acid, were more invasive than their normal cell counterparts, and that this was specifically reversed by adding exogenous TIMP-1 protein. The invasive cell population had characteristics of an early population of primitive mesenchymal cells, including expression of vimentin and a transient period of invasiveness from 4-8 d after initiation of differentiation. Therefore, metalloproteinase activity can be rate limiting for cell invasion.