Hotary K, Allen E, Punturieri A, Yana I, Weiss S J
Department of Internal Medicine and the University of Michigan Comprehensive Cancer Center, University of Michigan, Ann Arbor, Michigan 48109, USA.
J Cell Biol. 2000 Jun 12;149(6):1309-23. doi: 10.1083/jcb.149.6.1309.
During tissue-invasive events, migrating cells penetrate type I collagen-rich interstitial tissues by mobilizing undefined proteolytic enzymes. To screen for members of the matrix metalloproteinase (MMP) family that mediate collagen-invasive activity, an in vitro model system was developed wherein MDCK cells were stably transfected to overexpress each of ten different MMPs that have been linked to matrix remodeling states. MDCK cells were then stimulated with scatter factor/hepatocyte growth factor (SF/HGF) to initiate invasion and tubulogenesis atop either type I collagen or interstitial stroma to determine the ability of MMPs to accelerate, modify, or disrupt morphogenic responses. Neither secreted collagenases (MMP-1 and MMP-13), gelatinases (gelatinase A or B), stromelysins (MMP-3 and MMP-11), or matrilysin (MMP-7) affected SF/HGF-induced responses. By contrast, the membrane-anchored metalloproteinases, membrane-type 1 MMP, membrane-type 2 MMP, and membrane-type 3 MMP (MT1-, MT2-, and MT3-MMP) each modified the morphogenic program. Of the three MT-MMPs tested, only MT1-MMP and MT2-MMP were able to directly confer invasion-incompetent cells with the ability to penetrate type I collagen matrices. MT-MMP-dependent invasion proceeded independently of proMMP-2 activation, but required the enzymes to be membrane-anchored to the cell surface. These findings demonstrate that MT-MMP-expressing cells can penetrate and remodel type I collagen-rich tissues by using membrane-anchored metalloproteinases as pericellular collagenases.
在组织侵袭过程中，迁移细胞通过动员未知的蛋白水解酶穿透富含I型胶原蛋白的间质组织。为了筛选介导胶原蛋白侵袭活性的基质金属蛋白酶（MMP）家族成员，开发了一种体外模型系统，其中MDCK细胞被稳定转染以过表达与基质重塑状态相关的十种不同MMP中的每一种。然后用散射因子/肝细胞生长因子（SF/HGF）刺激MDCK细胞，以在I型胶原蛋白或间质基质上启动侵袭和管状发生，以确定MMP加速、改变或破坏形态发生反应的能力。分泌型胶原酶（MMP-1和MMP-13）、明胶酶（明胶酶A或B）、基质溶解素（MMP-3和MMP-11）或基质溶素（MMP-7）均未影响SF/HGF诱导的反应。相比之下，膜锚定金属蛋白酶，即膜型1 MMP、膜型2 MMP和膜型3 MMP（MT1-MMP、MT2-MMP和MT3-MMP）均改变了形态发生程序。在所测试的三种MT-MMP中，只有MT1-MMP和MT2-MMP能够直接赋予无侵袭能力的细胞穿透I型胶原基质的能力。MT-MMP依赖性侵袭独立于proMMP-2激活进行，但需要这些酶锚定在细胞表面。这些发现表明，表达MT-MMP的细胞可以通过使用膜锚定金属蛋白酶作为细胞周围胶原酶来穿透和重塑富含I型胶原蛋白的组织。
