MT1-MMP, but not secreted MMPs, influences the migration of human microvascular endothelial cells in 3-dimensional collagen gels
MT1-MMP, but not secreted MMPs, influences the migration of human microvascular endothelial cells in 3-dimensional collagen gels
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DOI：
10.1002/jcb.10257
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发表时间：
2002-01-01
影响因子：
4
通讯作者：
Reed, MJ
中科院分区：
文献类型：
--
作者：
Koike, T;Vernon, RB;Reed, MJ
基质金属蛋白酶(MMPs)及其特异性抑制物在血管生成中发挥重要作用。我们研究了人微血管内皮细胞(HmECs)在三维(3D)I型胶原凝胶和二维(2D)胶原涂层表面培养后，对特定MMPs和TIMPs表达的调节作用。经逆转录聚合酶链式反应(RT-PCR)检测，MMPs-1、MMP2、MMP13、MMP1-MMP1、TIMPS-1、TIMPS-2在2D和3D培养中的表达水平相似。通过Western印迹分析，TIMP-1和ProMMP1在2D和3D培养上清液中表达相似，而活性MMPs-1、-9和-13未检测到。细胞裂解物(CL)中存在活性的基质金属蛋白酶-13，且3D培养的裂解物中的活性高于2D培养的裂解液。与2D培养相比，CL和3D培养上清液中TIMP-2的表达减少，而ProMMP2和ProMT1-MMPs向活性或加工型转化增加。基质金属蛋白酶抑制剂GM6001干扰三维培养的人脐静脉内皮细胞的迁移，但不干扰二维培养的内皮细胞的迁移。加入活性的基质金属蛋白酶-1或阻断抗TIMP-1的抗体并不影响hmECs在3D胶原中的迁移。TIMP-2(MT1-MMPs的抑制剂)可减少3D胶原的迁移，但TIMP-1(Mt1-MMPs的抑制作用较差，但对MMP2的抑制作用较强)不影响其在3D胶原中的迁移。综上所述，我们的数据表明，MT1-MMPs对hmECs通过3D胶原的运动有显著贡献，而分泌型MMPs-1、-2、-9和-13对这一运动不是关键的。(C)2002年Wiley-Liss，Inc.
Matrix metalloproteinases (MMPs) and their specific inhibitors the TIMPs play significant roles in angiogenesis. We investigated how the expression of specific MMPs and TIMPs by human microvascular endothelial cells (hmECs) was modulated by culture of the cells in 3-dimensional (3D) type I collagen gels versus 2-dimensional (2D) collagen-coated surfaces. By reverse-transcription polymerase chain reaction (RT-PCR), levels of mRNA for MMPs-1, -2, and -13, MT1-MMP, and TIMPs-1 and -2 were similar in 2D versus 3D cultures. By Western blot assay, TIMP-1 and proMMP-1 were present and were expressed similarly in media from 2D versus 3D cultures, whereas active MMPs-1, -9, and -13 were not detected. Active MMP-13 was present in cell lysates (CL) and was increased in lysates from 3D cultures relative to 2D cultures. Relative to 2D cultures, CL and media from 3D cultures exhibited a decrease in expression of TIMP-2 and an increased conversion of proMMP-2 and proMT1-MMP to active or processed forms. The MMP inhibitor GM6001 interfered with the migration of hmECs in 3D cultures, but not in 2D cultures. Addition of active MMP-1 or blocking antibodies to TIMP-1 did not affect the migration of hmECs in 3D collagen. Migration in 3D collagen was decreased by TIMP-2 (an inhibitor of MT1-MMP), but not by TIMP-1 (a poor inhibitor of MT1-MMP, but an efficient inhibitor of MMP-2). Collectively, our data indicate that MT1-MMP contributes significantly to the movement of hmECs through 3D collagen, in contrast to secretory-type MMPs-1, -2, -9, and -13, which are not critical for this movement. (C) 2002 Wiley-Liss, Inc.