Granulocyte-macrophage colony stimulating factor induces endothelial capillary formation through induction of membrane-type 1 matrix metalloproteinase expression in vitro
Granulocyte-macrophage colony stimulating factor induces endothelial capillary formation through induction of membrane-type 1 matrix metalloproteinase expression in vitro
复制标题
DOI：
10.1002/ijc.23234
复制
发表时间：
2008-03-15
影响因子：
6.4
通讯作者：
English, William R.
中科院分区：
文献类型：
--
作者：
Krubasik, Davia;Eisenach, Patricia A.;English, William R.
在我们的研究中，我们使用体外模型研究了粒细胞巨噬细胞集落刺激因子（GM-CSF）调节血管生成的机制。 GM-CSF显着增加了接种在I型胶原凝胶中的内皮细胞球体的毛细血管前芽样形成以及内皮细胞与成纤维细胞共培养时的小管形成。在这两种情况下，芽和小管的形成高度依赖于金属蛋白酶活性。球体和共培养模型中的金属蛋白酶组织抑制剂 (TIMP) 分析显示 TIMP-2 具有抑制作用，但 TIMP-1 没有抑制作用，表明膜型基质金属蛋白酶 (MT-MMP) 的活性。发现 GM-CSF 诱导的球体中的芽形成可被 MT1-MMP 特异性的 siRNA 有效抑制。随后的分析表明，GM-CSF 以 MEK 依赖性机制短暂增加内皮细胞中 MT1-MMP mRNA，从而导致 MT1-MMP 表面水平增加。这伴随着 GMCSF 刺激的内皮细胞裂解物对 DQ 胶原的 MT1-MMP 依赖性降解的增加。 GM-CSF 不会增加成纤维细胞中 MT1-MMP 的水平。 GM-CSF对内皮细胞芽形成的影响可以通过用表达MT1-MMP的病毒对完整球体进行腺病毒转导来模拟，但不能通过在球体形成之前转导内皮细胞来模拟，这表明MT1-MMP的上调必定只发生在直接参与肾小管形成的细胞中。 (C) 2007 Wiley-Liss, Inc.
In our study, we examined the mechanism by which granulocyte-macrophage colony stimulating factor (GM-CSF) regulates angiogenesis using in vitro models. GM-CSF significantly increased precapillary sprout-like formation from endothelial cell spheroids seeded in type-I collagen gels and tubule formation on coculture of endothelial cells with fibroblasts. In both cases, sprout and tubule formation was highly dependent on metalloproteinase activity. Tissue Inhibitor of metalloproteinase (TIMP) profiling in the spheroid and coculture models showed inhibition by TIMP-2 but not by TIMP-1, indicative of activity of membrane-type matrix metalloproteinases (MT-MMPs). GM-CSF induced sprout formation in spheroids was found to be potently inhibited by siRNA specific for MT1-MMP. Subsequent analysis showed that GM-CSF transiently increased MT1-MMP mRNA in endothelial cells in a MEK-dependent mechanism, which led to increased surface levels of MT1-MMP. This was accompanied by an increase in MT1-MMP-dependent degradation of DQ-collagen by lysates of GMCSF stimulated endothelial cells. GM-CSF did not increase MT1-MMP levels in fibroblasts. The effect of GM-CSF on endothelial cell sprout formation could be mimicked by adenoviral transduction of intact spheroids with virus expressing MT1-MMP, but not by transduction of endothelial cells before spheroid formation, suggesting that upregulation of MT1-MMP must only occur in cells directly involved in tubule formation. (C) 2007 Wiley-Liss, Inc.