MT1-MMP Initiates Activation of pro-MMP-2 and Integrin αvβ3 Promotes Maturation of MMP-2 in Breast Carcinoma Cells

doi:10.1006/excr.2000.5118

Experimental Cell Research

Volume 263, Issue 2, 15 February 2001, Pages 209-223

https://doi.org/10.1006/excr.2000.5118 Get rights and content

Abstract

We evaluated cellular mechanisms involved in the activation pathway of matrix prometalloproteinase-2 (pro-MMP-2), an enzyme implicated in the malignant progression of many tumor types. Membrane type-1 matrix metalloproteinase (MT1-MMP) cleaves the N-terminal prodomain of pro-MMP-2 thus generating the activation intermediate that then matures into the fully active enzyme of MMP-2. Our results provide evidence on how a collaboration between MT1-MMP and integrin αvβ3 promotes more efficient activation and specific, transient docking of the activation intermediate and, further, the mature, active enzyme of MMP-2 at discrete regions of cells. We show that coexpression of MT1-MMP and integrin αvβ3 in MCF7 breast carcinoma cells specifically enhances in trans autocatalytic maturation of MMP-2. The association of MMP-2′s C-terminal hemopexin-like domain with those molecules of integrin αvβ3 which are proximal to MT1-MMP facilitates MMP-2 maturation. Vitronectin, a specific ligand of integrin αvβ3, competitively blocked the integrin-dependent maturation of MMP-2. Immunofluorescence and immunoprecipitation studies supported clustering of MT1-MMP and integrin αvβ3 at discrete regions of the cell surface. Evidently, the identified mechanisms appear to be instrumental to clustering active MMP-2 directly at the invadopodia and invasive front of αvβ3-expressing cells or in their close vicinity, thereby accelerating tumor cell locomotion.

References (44)

L.M. Coussens et al.
Matrix metalloproteinases and the development of cancer
Chem. Biol.
(1996)
E.C. Kohn et al.
Molecular insights into cancer invasion: Strategies for prevention and intervention
Cancer Res.
(1995)
H. Nagase et al.
Matrix metalloproteinases
J. Biol. Chem.
(1999)
H. Sato et al.
A matrix metalloproteinase expressed on the surface of invasive tumour cells
Nature
(1994)
A.Y. Strongin et al.
Mechanism of cell surface activation of 72-kDa type IV collagenase. Isolation of the activated form of the membrane metalloprotease
J. Biol. Chem.
(1995)
A. Okada et al.
Membrane-type matrix metalloproteinase (MT-MMP) gene is expressed in stromal cells of human colon, breast, and head and neck carcinomas
Proc. Natl. Acad. Sci. USA
(1995)
A.Y. Strongin et al.
Plasma membrane-dependent activation of the 72-kDa type IV collagenase is prevented by complex formation with TIMP-2
J. Biol. Chem.
(1993)
H. Nakamura et al.
Enhanced production and activation of progelatinase A mediated by membrane-type 1 matrix metalloproteinase in human papillary thyroid carcinomas
Cancer Res.
(1999)
P.C. Brooks et al.
Localization of matrix metalloproteinase MMP-2 to the surface of invasive cells by interaction with integrin alpha v beta 3
Cell
(1996)
W.L. Monsky et al.
Binding and localization of M(r) 72,000 matrix metalloproteinase at cell surface invadopodia
Cancer Res.
(1993)

C.A. Partridge et al.

Localization and activation of type IV collagenase/gelatinase at endothelial focal contacts

Am. J. Physiol.

(1997)

E.I. Deryugina et al.

Matrix metalloproteinase-2 activation modulates glioma cell migration

J. Cell Sci.

(1997)

P.C. Brooks et al.

Disruption of angiogenesis by PEX, a noncatalytic metalloproteinase fragment with integrin binding activity

Cell

(1998)

M.I. Cockett et al.

Matrix metalloproteinases and metastatic cancer

Biochem. Soc. Symp.

(1998)

T. Itoh et al.

Reduced angiogenesis and tumor progression in gelatinase A-deficient mice

Cancer Res.

(1998)

M. Yamamoto et al.

Differential expression of membrane-type matrix metalloproteinase and its correlation with gelatinase A activation in human malignant brain tumors in vivo and in vitro

Cancer Res.

(1996)

G.I. Goldberg et al.

Human 72-kilodalton type IV collagenase forms a complex with a tissue inhibitor of metalloproteases designated TIMP-2

Proc. Natl. Acad. Sci. USA

(1989)

Toth M. Hernandez-Barrantes et al.

Binding of active (57 kDa) membrane type 1-matrix metalloproteinase (MT1-MMP) to tissue inhibitor of metalloproteinase (TIMP)-2 regulates MT1-MMP processing and pro-MMP-2 activation

J. Biol. Chem.

(2000)

J.J. Caterina et al.

Inactivating mutation of the mouse tissue inhibitor of metalloproteinases-2 (TIMP-2) gene alters proMMP-2 activation

J. Biol. Chem.

(2000)

Z. Wang et al.

TIMP-2 is required for efficient activation of proMMP-2 in vivo

J. Biol. Chem.

(2000)

E.I. Deryugina et al.

Remodeling of collagen matrix by human tumor cells requires activation and cell surface association of matrix metalloproteinase-2

Cancer Res.

(1998)

Cited by (336)

Co-treatment of Nimbolide augmented the anti-arthritic effects of methotrexate while protecting against organ toxicities
2022, Life Sciences
Prolonged exposure to the pharmacological doses of disease-modifying anti-rheumatic drugs (DMARDs) often results in major organ toxicities resulting in poor patient compliance. Methotrexate (MTX) is one of the commonly prescribed DMARDs for the treatment of arthritis, which results in vital organ dysfunction. To retain the anti-arthritic activity of MTX with the reduction in toxicities, combination therapies are warranted. Nimbolide (NMB) is a potent anticancer, anti-inflammatory and anti-fibrotic agent whose potential has been demonstrated in various pre-clinical models. Monoarthritis was developed with Complete Freund's Adjuvant in the knees of Wistar rats and treatment was given with either NMB (3 mg/kg/day) or MTX (2 mg/kg/week) alone or combination therapy (NMB + MTX). The anti-arthritic effects were evaluated by arthritic scoring, radiological imaging, synovial tissue proteins analysis, and histopathological staining. While hepato-renal toxicity was assessed in serum by evaluating the kidney and liver functional parameters, in tissues by oxidative-nitrosative stress markers, and pro-inflammatory cytokines levels. Histopathological analysis was performed to study the extent of tissue damage. Molecular studies like immunoblotting and immunohistochemistry were performed to understand the effect of combination therapy. We thereby report that monotherapy with either NMB or MTX exhibited significant anti-arthritic effects, while combination therapy resulted in augmented anti-arthritic effects with significant reduction in hepato-renal toxicity produced by MTX probably through anti-inflammatory and anti-oxidant effects. Therefore, our proposed combination of NMB and MTX may serve as a potential strategy for the effective management of arthritis.
Biosensors for Caspase-3: From chemical methodologies to biomedical applications
2022, Talanta
Caspase-3 plays irreplaceable roles in apoptosis and related diseases. An imbalance in the measured levels of Caspase-3 is implicated in irreversible apoptosis. Therefore, the detection of Caspase-3 is of great significance for apoptosis imaging and the evaluation effect of early tumor treatment and other diseases. Herein, advances in the recent innovations of Caspase-3 response fluorescence biosensors, including molecular probes and nanoprobes, are systematically summarized in sections corresponding. The performances of various luminescence probes in Caspase-3 detection are discussed intensively in the design strategy of chemical structure, response mechanism and biological application. Finally, the current challenges and prospects of the design of new Caspase-3 responsive fluorescence probes for apoptosis imaging, or similar molecular event are proposed.
Inhibition of β1 integrin induces its association with MT1-MMP and decreases MT1-MMP internalization and cellular invasiveness
2021, Cellular Signalling
Citation Excerpt :
Consistent with these observations, microscopic analyses revealed increased levels of both β1 integrin and MT1-MMP on the cell surface of non-permeabilized, β1-integrin-inhibited cells, at both 3 h and 16 h timepoints (Fig. 3E). It is well recognized that clustering of integrin receptors plays a role in modulating downstream signaling events, and that integrin clustering occurs at invadopodia [9,30,43]. Previous studies have also revealed that clustering of β1 integrin receptors with MT1-MMP functions to stabilize their interaction and leads to a subsequent increase in their surface expression [11].
Integrin signaling plays a fundamental role in the establishment of focal adhesions and the subsequent formation of invadopodia in malignant cancer cells. Invadopodia facilitate localized adhesion and degradation of the extracellular matrix (ECM), which promote tumour cell invasion and metastasis. Degradation of ECM components is often driven by membrane type-1 matrix metalloproteinase (MT1-MMP), and we have recently shown that regulation of enzyme internalization is dependent on signaling downstream of β1 integrin. Phosphorylation of the cytoplasmic tail of MT1-MMP is required for its internalization and delivery to Rab5-marked early endosomes, where it is then able to be recycled to new sites of invadopodia formation and promote invasion. Here we found that inhibition of β1 integrin, using the antibody AIIB2, inhibited the internalization and recycling of MT1-MMP that is necessary to support long-term cellular invasion. MT1-MMP and β1 integrin were sequestered at the cell surface when β1-integrin was inhibited, and their association under these conditions was detected using immunoprecipitation and mass spectrometry analyses. Sequestration of β1 integrin and MT1-MMP at the cell surface resulted in the formation of large invadopodia and local ECM degradation; however, the impaired internalization and recycling of MT1-MMP and β1 integrin ultimately led to a loss of invasive behaviour.
Nimbolide exerts protective effects in complete Freund's adjuvant induced inflammatory arthritis via abrogation of STAT-3/NF-κB/Notch-1 signaling
2021, Life Sciences
Activation of transmembrane Notch-1 receptors through inflammatory cytokines is highly regulated by STAT-3 and NF-κB phosphorylation. Nimbolide (NMB) exhibits potent anti-inflammatory, anti-fibrotic, anticancer activities by targeting various pathways. Here, we have investigated the effect of NMB in regulation of STAT-3/NF-κB/Notch-1 axis in complete Freund's adjuvant (CFA) induced inflammatory arthritis (IA) model.
The anti-inflammatory and anti-arthritic activity of NMB was evaluated both in vitro (IL-1β stimulated HIG-82 synovial fibroblasts) and in vivo (CFA induced rat model of IA) models. In vitro anti-arthritic activity was assessed by anti-migratory effect, while in vivo effects were evaluated through radiological and histological analysis. The effect of NMB on STAT-3, NF-κB, Notch-1 signaling pathways and proinflammatory cytokines were studied using western blot, immunohistochemistry and ELISA methods. Key findings NMB attenuated the migration of synovial fibroblasts in vitro. It reduced the progression of arthritis as evidenced from the improved radiological and histological abnormalities in arthritic rats. NMB significantly suppressed the nitrosooxidative stress and levels of pro-inflammatory cytokines. NMB also exhibited remarkable protective activity against upregulation of MAPK, STAT-3 and NF-κB phosphorylation mediated Notch-1 signaling pathway in synovial tissue of arthritic rats.
NMB may have clinical therapeutic value in rheumatoid arthritis by inhibiting STAT-3/NF-κB/Notch-1 axis and also by reducing the levels of proinflammatory cytokines.
The multiple roles of actin-binding proteins at invadopodia
2021, International Review of Cell and Molecular Biology
Invadopodia are actin-rich membrane protrusions that facilitate cancer cell dissemination by focusing on proteolytic activity and clearing paths for migration through physical barriers, such as basement membranes, dense extracellular matrices, and endothelial cell junctions. Invadopodium formation and activity require spatially and temporally regulated changes in actin filament organization and dynamics. About three decades of research have led to a remarkable understanding of how these changes are orchestrated by sequential recruitment and coordinated activity of different sets of actin-binding proteins. In this chapter, we provide an update on the roles of the actin cytoskeleton during the main stages of invadopodium development with a particular focus on actin polymerization machineries and production of pushing forces driving extracellular matrix remodeling.
Recent advances in the development of activatable multifunctional probes for in vivo imaging of caspase-3
2021, Chinese Chemical Letters
Caspases are a family of proteases that play critical roles in controlling inflammation and cell death. Apoptosis is a caspase-3 mainly controlled behavior to avoid inflammation and damage to surrounding cells, whereas anomalistic cell apoptosis may be associated with many diseases. The detection and imaging of caspase-3 will be of great significance in evaluating the early therapeutic effect of tumors. Developing smart fluorescent probes may be helpful for the visualization of therapeutic effect compared with “always on” probes. Thus, more and more works toward activatable fluorescent probes for caspase-3 imaging have been reported. In addition, multifunctional probes have also been designed to further improve the imaging of caspase-3. Herein, this review systematically summarized the representative work of caspase-3 from the perspective of molecular design that it will play a guiding role in the design of probes that respond to caspase-3. Also, challenges and perspectives toward the field for imaging of cell apoptosis (caspase-3) are also discussed.

View all citing articles on Scopus

¹: To whom correspondence and reprint requests should be addressed at The Burnham Institute, 10901 North Torrey Pines Road, La Jolla, CA 92037. Fax: (858) 646-3192. E-mail: [email protected].

View full text

Regular ArticleMT1-MMP Initiates Activation of pro-MMP-2 and Integrin αvβ3 Promotes Maturation of MMP-2 in Breast Carcinoma Cells

Abstract

Matrix metalloproteinases and the development of cancer

Chem. Biol.

Molecular insights into cancer invasion: Strategies for prevention and intervention

Cancer Res.

Matrix metalloproteinases

J. Biol. Chem.

A matrix metalloproteinase expressed on the surface of invasive tumour cells

Nature

Mechanism of cell surface activation of 72-kDa type IV collagenase. Isolation of the activated form of the membrane metalloprotease

J. Biol. Chem.

Membrane-type matrix metalloproteinase (MT-MMP) gene is expressed in stromal cells of human colon, breast, and head and neck carcinomas

Proc. Natl. Acad. Sci. USA

Plasma membrane-dependent activation of the 72-kDa type IV collagenase is prevented by complex formation with TIMP-2

J. Biol. Chem.

Enhanced production and activation of progelatinase A mediated by membrane-type 1 matrix metalloproteinase in human papillary thyroid carcinomas

Cancer Res.

Localization of matrix metalloproteinase MMP-2 to the surface of invasive cells by interaction with integrin alpha v beta 3

Cell

Binding and localization of M(r) 72,000 matrix metalloproteinase at cell surface invadopodia

Cancer Res.

Localization and activation of type IV collagenase/gelatinase at endothelial focal contacts

Am. J. Physiol.

Matrix metalloproteinase-2 activation modulates glioma cell migration

J. Cell Sci.

Disruption of angiogenesis by PEX, a noncatalytic metalloproteinase fragment with integrin binding activity

Cell

Matrix metalloproteinases and metastatic cancer

Biochem. Soc. Symp.

Reduced angiogenesis and tumor progression in gelatinase A-deficient mice

Cancer Res.

Differential expression of membrane-type matrix metalloproteinase and its correlation with gelatinase A activation in human malignant brain tumors in vivo and in vitro

Cancer Res.

Human 72-kilodalton type IV collagenase forms a complex with a tissue inhibitor of metalloproteases designated TIMP-2

Proc. Natl. Acad. Sci. USA

Binding of active (57 kDa) membrane type 1-matrix metalloproteinase (MT1-MMP) to tissue inhibitor of metalloproteinase (TIMP)-2 regulates MT1-MMP processing and pro-MMP-2 activation

J. Biol. Chem.

Inactivating mutation of the mouse tissue inhibitor of metalloproteinases-2 (TIMP-2) gene alters proMMP-2 activation

J. Biol. Chem.

TIMP-2 is required for efficient activation of proMMP-2 in vivo

J. Biol. Chem.

Remodeling of collagen matrix by human tumor cells requires activation and cell surface association of matrix metalloproteinase-2

Cancer Res.

Regular Article
MT1-MMP Initiates Activation of pro-MMP-2 and Integrin αvβ3 Promotes Maturation of MMP-2 in Breast Carcinoma Cells