Shankavaram U T, Lai W C, Netzel-Arnett S, Mangan P R, Ardans J A, Caterina N, Stetler-Stevenson W G, Birkedal-Hansen H, Wahl L M
Immunopathology Section and Matrix Metalloproteinase Unit, National Institute of Dental and Craniofacial Research and the Extracellular Matrix Pathology Section, Laboratory of Pathology, NCI, National Institutes of Health, Bethesda, Maryland 20892, USA.
J Biol Chem. 2001 Jun 1;276(22):19027-32. doi: 10.1074/jbc.M009562200. Epub 2001 Mar 20.
Membrane type 1-matrix metalloproteinase (MT1-MMP)-mediated activation of MMP-2 is thought to be important in the proteolysis of extracellular matrix in pathological events in which monocytes/macrophages are found. Here we report on the induction and regulation of human monocyte MT1-MMP and its role in MMP-2 activation. Activation of monocytes by lipopolysaccharide resulted in the induction of MT1-MMP mRNA and protein that was suppressed by inhibitors of prostaglandin synthesis (indomethacin), adenylyl cyclase (SQ 22536), and protein kinase A (Rp-cAMPs). Suppression of MT1-MMP by indomethacin and SQ 22536 was reversed by prostaglandin E(2) and dibutyryl cyclic AMP, respectively, demonstrating that induction of monocyte MT1-MMP is regulated through a prostaglandin-cAMP pathway. Functional analysis revealed that pro-MMP-2 in the supernatants from human bone marrow stromal fibroblasts, normal male-derived fibroblasts and melanoma cells (A2058) was converted to active MMP-2 when cultured with activated but not control monocytes. Antibodies against MT1-MMP blocked the activation of MMP-2. Tissue inhibitor of metalloproteinase-2 regulation of MMP-2 activation was shown through the addition of varying amounts of recombinant tissue inhibitor of metalloproteinase-2 with pro-MMP-2 to MT1-MMP-expressing monocytes. These findings demonstrate that activated monocytes express functionally active MT1-MMP that may play a significant role in the activation of MMP-2 produced by other cells and as such influence developmental and pathological conditions.
膜型1-基质金属蛋白酶（MT1-MMP）介导的MMP-2激活被认为在有单核细胞/巨噬细胞参与的病理事件中细胞外基质的蛋白水解过程中起重要作用。在此，我们报告人单核细胞MT1-MMP的诱导、调控及其在MMP-2激活中的作用。脂多糖激活单核细胞导致MT1-MMP mRNA和蛋白的诱导，这被前列腺素合成抑制剂（吲哚美辛）、腺苷酸环化酶抑制剂（SQ 22536）和蛋白激酶A抑制剂（Rp-cAMPs）所抑制。吲哚美辛和SQ 22536对MT1-MMP的抑制分别被前列腺素E2和二丁酰环磷酸腺苷逆转，表明单核细胞MT1-MMP的诱导是通过前列腺素-cAMP途径调控的。功能分析显示，当与人骨髓基质成纤维细胞、正常男性来源的成纤维细胞和黑色素瘤细胞（A2058）的上清液中的前MMP-2与活化而非对照单核细胞共培养时，前MMP-2转化为活性MMP-2。抗MT1-MMP抗体阻断了MMP-2的激活。通过向表达MT1-MMP的单核细胞中加入不同量的重组金属蛋白酶组织抑制剂-（TIMP-2）与前MMP-2，显示了TIMP-2对MMP-2激活的调节作用。这些发现表明，活化的单核细胞表达具有功能活性的MT1-MMP，其可能在激活其他细胞产生的MMP-2中起重要作用，从而影响发育和病理状况。
