Membrane Type 1-Matrix Metalloproteinase Is Activated during Migration of Human Endothelial Cells and Modulates Endothelial Motility and Matrix Remodeling*
- Beatriz G. Gálvez‡§,
- Salomón Matı́as-Román‡,
- Juan P. Albar¶,
- Francisco Sánchez-Madrid‡ and
- Alicia G. Arroyo‡‖
- From the ‡Departamento de Inmunologı́a, Hospital de la Princesa, 28006 Madrid and the ¶Immunology and Oncology Department, Pharmacia-Consejo Superior de Investigaciones Cientı́ficas, Centro Nacional de Biotecnologı́a, 28049 Madrid, Spain
Abstract
Matrix metalloproteinases are thought to play an important role in endothelial cell migration and matrix remodeling. We have used an in vitro wound healing migration model and newly generated anti-membrane type 1-matrix metalloproteinase (MT1-MMP) monoclonal antibodies (mAbs) to characterize the role of MT1-MMP during this process. First, the expression and shedding of MT1-MMP are up-regulated upon induction of migration in endothelial cells, as demonstrated by flow cytometry and Western blot analysis. Furthermore, MT1-MMP is concentrated at discrete areas in migrating endothelial cells, in contrast to the diffuse pattern observed in confluent cells. Interestingly, migration of endothelial cells results in the stimulation of MT1-MMP activity, as shown by its ability to process pro-MMP-2 and to degrade fibrinogen assessed by zymography. Moreover, MT1-MMP-mediated gelatin degradation is enriched at migration sites. mAbs generated against the MT1-MMP catalytic domain are shown to inhibit MT1-MMP enzymatic activity and to impair both phorbol 12-myristate 13-acetate-induced endothelial migration and invasion of collagen and fibrin gels. Furthermore, a reduction in the formation of capillary tubes in Matrigel is also observed when endothelial cells are pretreated with the blocking anti-MT1-MMP mAbs. Altogether, these data demonstrate that MT1-MMP plays an important role during endothelial cell migration, and its activity can modulate endothelial migration, invasion, and formation of capillary tubes during the angiogenic response.
Footnotes
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↵* This work was supported in part by Grants FIS00/0114 from Fondo de Investigaciones Sanitarias and CAM 08.3/0003.1/2000 from Comunidad Autónoma de Madrid (to A. G. A.).The costs of publication of this article were defrayed in part by the payment of page charges. The article must therefore be hereby marked “advertisement” in accordance with 18 U.S.C. Section 1734 solely to indicate this fact.
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↵§ Recipient of a predoctoral fellowship from the Comunidad Autónoma de Madrid.
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↵‖ Recipient of a grant from Fondo de Investigaciones Sanitarias. To whom correspondence should be addressed: Dept. de Inmunologı́a. Hospital de la Princesa, C/Diego de León 62, 28006 Madrid, Spain. Tel.: 34-91-5202334; Fax: 34-91-5202374; E-mail: agarciaa@hlpr.insalud.es.
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Published, JBC Papers in Press, July 11, 2001, DOI 10.1074/jbc.M104094200
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↵2 B. G. Gálvez and A. G. Arroyo, unpublished observations.
- Abbreviations:
- MMP
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matrix metalloproteinase
- ECM
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extracellular matrix
- HUVEC
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human endothelial cell(s) from umbilical vein
- MT1-MMP
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membrane type 1-matrix metalloproteinase
- mAb
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monoclonal antibody
- PBS
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phosphate-buffered saline
- FITC
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fluorescein isothiocyanate
- PAGE
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polyacrylamide gel electrophoresis
- PMA
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phorbol 12-myristate 13-acetate
- MFI
-
mean fluorescence intensity
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- Received May 7, 2001.
- Revision received June 15, 2001.
- The American Society for Biochemistry and Molecular Biology, Inc.
