Carbohydrate-Mediated Regulation of Matrix Metalloproteinase-2 Activation in Normal Human Fibroblasts and Fibrosarcoma Cells

Gervasi, David C.; Raz, Avraham; Dehem, Marie; Yang, Maozhou; Kurkinen, Markku; Fridman, Rafael

doi:10.1006/bbrc.1996.1694

Cited by 39 publications

(24 citation statements)

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“…In agreement with this hypothesis, it is worth noting that Stanton and coworkers reported that HT1080 cells treated with phor-bol ester processed pro-MMP-2 into the 59-kDa fully mature form. In our cell culture model, and in agreement with others [56], a similar treatment promoted the generation of the 62-kDa intermediate MMP-2 species without any significant further conversion into the mature form.…”

Section: Type IV Collagen Induces the Formation Of Mature 59-kda Mmp-2supporting

confidence: 92%

“…As previously reported by others [6,13,15,[56][57][58], we showed that TPA-or ConA-treated HT1080 cells increased MT1-MMP mRNA expression and substantially upregulated MT1-MMP protein synthesis. Furthermore, both treatments induced the formation of a 43-kDa MT1-MMP species (Figs.…”

Section: Mmp-2 Activation By Tpa-and Cona-treated Ht1080 Cellssupporting

confidence: 89%

“…6B, lanes 2-4). In this respect, it is interesting to note that even if both TPA and ConA treatments induced the formation of the 62-kDa intermediate MMP-2, only ConA efficiently converted this intermediate into the mature 59-kDa form, as previously reported by Gervasi and coworkers [56]. The ability of ConA to promote the second step of MMP-2 activation (i.e., the intermolecular autolytic cleavage) has been ascribed, at least in part, to its potential to cluster membrane glycoproteins, including MT1-MMP, thereby favoring the autoproteolytic processing of adjacent MT1-MMP-associated intermediate MMP-2 species [56,60,61].…”

Section: Mmp-2 Activation By Tpa-and Cona-treated Ht1080 Cellssupporting

confidence: 76%

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Type IV Collagen Induces Matrix Metalloproteinase 2 Activation in HT1080 Fibrosarcoma Cells

Maquoi

Frankenne

Noël

et al. 2000

Experimental Cell Research

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Matrix metalloproteinase 2 (MMP-2) activation has been described as a "master switch" which triggers tumor spread and metastatic progression. We show here that type IV collagen, a major component of basement membranes, promotes MMP-2 activation by HT1080 cells. When plated on plastic, HT1080 cells constitutively processed the 66-kDa pro-MMP-2 into a 62-kDa intermediate activated form, most probably through a membrane type (MT) 1 MMP-dependent mechanism. In the presence of type IV collagen, part of this intermediate form was further processed to fully activated 59-kDa MMP-2. This activation was prevented by tissue inhibitor of MMP (TIMP)-2 and a broad-spectrum hydroxamic acid-based synthetic MMP inhibitor (GI129471). Type IV collagen-mediated pro-MMP-2 activation did not involve either a transcriptional modulation of MMP-2, MT1-MMP, or TIMP-2 expression nor any alteration of MT1-MMP protein synthesis or processing. An inverse relationship between MMP-2 activation and the concentration of secreted TIMP-2 was observed. This is consistent with our previous report that TIMP-2 degradation is probably linked to the MT1-MMP-dependent MMP-2 activation mechanism. Because invasive tumor cells must breach basement membranes at different steps of the metastatic dissemination, the ability of HT1080 cells to activate pro-MMP-2 in the presence of type IV collagen might represent a key regulatory mechanism for the acquisition of an invasive potential.

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Section: Type IV Collagen Induces the Formation Of Mature 59-kda Mmp-2supporting

confidence: 92%

Section: Mmp-2 Activation By Tpa-and Cona-treated Ht1080 Cellssupporting

confidence: 89%

Section: Mmp-2 Activation By Tpa-and Cona-treated Ht1080 Cellssupporting

confidence: 76%

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Type IV Collagen Induces Matrix Metalloproteinase 2 Activation in HT1080 Fibrosarcoma Cells

Maquoi

Frankenne

Noël

et al. 2000

Experimental Cell Research

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“…Cell Culture-The characteristics and culture conditions of nonmalignant monkey kidney epithelial BS-C-1 (CCL-26) (28), human fibrosarcoma HT-1080 (CCL-121) (29), human fibroblasts HFL1 (CCL-153) (29), and human breast carcinoma MDA-MB-231 (HTB-26) (30) cells have been previously described. Human glioblastoma U-87 MG (HTB-14) cells, obtained from the American Tissue Culture Collection (ATCC), and immortalized homozygous Timp2 (Ϫ/Ϫ) mutant mouse fibroblasts, a gift from Dr. P. Soloway (Roswell Park Cancer Institute, Buffalo, NY) (20,31), were maintained in Dulbecco's modified Eagle's medium (DMEM) supplemented with 10% fetal bovine serum and antibiotics.…”

Section: Methodsmentioning

confidence: 99%

Complex Pattern of Membrane Type 1 Matrix Metalloproteinase Shedding

Tóth¹,

Hernandez-Barrantes²,

Osenkowski³

et al. 2002

Journal of Biological Chemistry

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112

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Membrane type 1 matrix metalloproteinase (MT1-MMP 255 and Ile 256 residues near the conserved methionine turn, a structural feature of the catalytic domain of all MMPs. Consistently, a recombinant 18-kDa fragment had no catalytic activity and did not bind TIMP-2. Thus, autocatalytic shedding evolved as a specific mechanism to terminate MT1-MMP activity on the cell surface by disrupting enzyme integrity at a vital structural site. In contrast, functional data suggest that the non-autocatalytic shedding generates soluble active MT1-MMP species capable of binding TIMP-2. These studies suggest that ectodomain shedding regulates the pericellular and extracellular activities of MT1-MMP through a delicate balance of active and inactive enzyme-soluble fragments.Release of the extracellular portion of type I transmembrane proteins, referred to as ectodomain shedding, has been established as a major regulatory mechanism to control the activity of a variety of membrane-bound proteins on the cell surface (1). Recent evidence suggests that ectodomain shedding is also characteristic of the membrane type matrix metalloproteinases (MT-MMPs), 1 a subfamily of membrane-anchored MMPs by means of a transmembrane domain or a glycosylphosphatidylinositol anchor (2, 3). The MT-MMPs are major mediators of proteolytic events on the cell surface, including turnover of extracellular matrix components (4, 5), cleavage of various surface adhesion receptors (6 -8), and initiation of zymogen activation cascades (9, 10). Uncontrolled MT-MMP activity contributes to abnormal development (11) and is a key determinant in cancer metastasis and tumor angiogenesis (12)(13)(14). To control the extent of pericellular activity, the MT-MMPs are inhibited by the tissue inhibitors of metalloproteinases (TIMPs), a family of natural protein MMP inhibitors. In addition, MT-MMPs have a unique regulatory mechanism in which the active enzyme undergoes a series of processing steps, either autocatalytic (15-17) or mediated by other proteases (18), that regulate the activity and nature of the enzyme species at the cell surface and at the pericellular space. Previous studies have shown that active MT1-MMP is autocatalytically processed on the cell surface to an inactive membrane-tethered ϳ44-kDa species lacking the entire catalytic domain (17). This processing is inhibited by TIMP-2, TIMP-4, and synthetic MMP inhibitors consistent with being an intermolecular autocatalytic event (19,20). Inhibition of MT1-MMP processing induces accumulation of the active enzyme on the cell surface, and, as a consequence, net MT1-MMP-dependent proteolysis is enhanced. Indeed, we have shown that, under certain conditions, inhibition of MT1-MMP autocatalysis by synthetic MMP inhibitors enhances pro-MMP-2 activation by MT1-MMP in the presence of TIMP-2 (20, 21). Thus, although the presence of inhibitors will stabilize MT1-MMP on the cell surface, the absence or reduced levels of inhibitors will facilitate autocatalysis. As a membrane-anchored protein, the autocatalytic processing of ac...

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“…Prior to the migration assay, the cells were treated with serum-free medium containing mitomycin C (25 g/ml,), in the presence and absence of concanavalin A (25 g/ml, 30 min) to induce pro-MMP-2 activation (20). Scratch wounds were then carefully made in the confluent monolayer using a disposable plastic pipette tip.…”

Section: Volume 280 • Number 40 • October 7 2005mentioning

confidence: 99%

Potent Mechanism-based Inhibitors for Matrix Metalloproteinases

Ikejiri

Bernardo

Bonfil

et al. 2005

Journal of Biological Chemistry

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117

118

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Matrix metalloproteinases (MMPs) are zinc-dependent endopeptidases that play important roles in physiological and pathological conditions. Both gelatinases (MMP-2 and -9) and membranetype 1 MMP (MMP-14) are important targets for inhibition, since their roles in various diseases, including cancer, have been well established. We describe herein a set of mechanism-based inhibitors that show high selectivity to gelatinases and MMP-14 (inhibitor 3) and to only MMP-2 (inhibitors 5 and 7). These molecules bind to the active sites of these enzymes, initiating a slow binding profile for the onset of inhibition, which leads to covalent enzyme modification. The full kinetic analysis for the inhibitors is reported. These are nanomolar inhibitors (K i ) for the formation of the noncovalent enzyme-inhibitor complexes. The onset of slow binding inhibition is rapid (k on of 10 2 to 10 4 M ؊1 s ؊1 ), and the reversal of the process is slow (k off of 10 ؊3 to 10 ؊4 s ؊1 ). However, with the onset of covalent chemistry with the best of these inhibitors (e.g. inhibitor 3), very little recovery of activity (<10%) was seen over 48 h of dialysis. We previously reported that broad spectrum MMP inhibitors like GM6001 enhance MT1-MMP-dependent activation of pro-MMP-2 in the presence of tissue inhibitor of metalloproteinases-2. Herein, we show that inhibitor 3, in contrast to GM6001, had no effect on pro-MMP-2 activation by MT1-MMP. Furthermore, inhibitor 3 reduced tumor cell migration and invasion in vitro. These results show that these new inhibitors are promising candidates for selective inhibition of MMPs in animal models of relevant human diseases.Extracellular proteolysis is an essential aspect of both physiological and pathological processes. Several enzyme families have been implicated in extracellular proteolysis, of which the matrix metalloproteinases (MMPs) 3 constitute an important group. The MMPs are zinc-dependent endopeptidases that play key roles in embryonic development, neurological processes, wound healing, angiogenesis, arthritis, cardiovascular diseases, and cancer, just to mention a few examples. In cancer, for instance, MMPs are implicated at all stages of tumor progression, including tumor growth, angiogenesis, and metastasis (1). Two MMPs, gelatinases A and B (MMP-2 and MMP-9, respectively), are highly expressed in human cancer, and a direct relationship between cancer progression and gelatinase expression and activity has been well established in many studies (2). As tumors manifest high levels of gelatinase activity, inhibitors specific for the gelatinases are highly sought.In the past 8 years, there have been numerous approaches aimed at targeting MMP activities in tumors, and several clinical trials were carried out to test the efficacy of various inhibitors. Unfortunately, the results of these trials were disappointing due to the lack of an objective clinical response and undesired side effects. Many reasons have been postulated for these effects, but at the core of the problem remains the issue of inhibitor ...

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Carbohydrate-Mediated Regulation of Matrix Metalloproteinase-2 Activation in Normal Human Fibroblasts and Fibrosarcoma Cells

Cited by 39 publications

References 0 publications

Type IV Collagen Induces Matrix Metalloproteinase 2 Activation in HT1080 Fibrosarcoma Cells

Type IV Collagen Induces Matrix Metalloproteinase 2 Activation in HT1080 Fibrosarcoma Cells

Complex Pattern of Membrane Type 1 Matrix Metalloproteinase Shedding

Potent Mechanism-based Inhibitors for Matrix Metalloproteinases

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