Elsevier

Matrix Biology

Volume 17, Issue 4, August 1998, Pages 293-304
Matrix Biology

Regular paper
Collagenase-1, stromelysin-1 and 92 kDa gelatinase are associated with tumor necrosis factor-α induced morphological change of human endothelial cells in Vitro

https://doi.org/10.1016/S0945-053X(98)90082-8Get rights and content

Abstract

Recently, we have shown that the tumor necrosis factor-α (TNF-α)-induced morphological change of EA.hy 926 human endothelial cells is associated with a decrease in the net synthesis of two proteoglycans (PGs), biglycan and syndecan-1, both of which have been suggested to play a role in cell adhesion. Here we have examined whether this phenotypic modulation of EA.hy 926 cells also involves altered expression of matrix metalloproteinases (MMPs) or their tissue inhibitors (TIMPs). We demonstrate that, when forming cobblestone-like monolayer cultures, these cells express and synthesize collagenase-1 (MMP-1), stromelysin-1 (MMP-3) and 72 kDa (MMP-2) and 92 kDa (MMP-9) gelatinases, all of which have previously been found in either normal or pathological human vascular wall. EA.hy 926 cells also express membrane-type1 MMP (MT1-MMP), but not matrilysin (MMP-7) and collagenase-3 (MMP-13). As regards TIMPs, we show that these cells express TIMP-1 and TIMP-2, but not TIMP-3 or TIMP-4. Exposure of the cells to TNF-α changed the cell morphology from a polygonal shape into a spindle shape and also increased the mRNA levels of MMP-1, MMP-3 and MMP-9, but slightly decreased the MMP-2 mRNA level. No change at the mRNA level of MT1-MMP was observed. Similarly to unstimulated cultures, no mRNA for MMP-7 or MMP-13 was detected in the TNF-α treated cultures. TNF-α had no effect on the TIMP-1 and TIMP-2 mRNA levels and did not induce TIMP-3 or TIMP-4 expression. Gelatin zymography and Western blot analysis revealed that the increase observed at the mRNA level of MMP-3 and MMP-9 was similar to that of their net protein level; furthermore, the active form of MMP-1 was induced. Our results indicate that the TNF-α-induced morphological change of EA.hy 926 cells is associated not only with specific changes in the expression of PGs by the cells, but also with specific changes in the expression of MMPs.

References (67)

  • H.T. Järveläinen et al.

    Differential expression of small chondroitin/dermatan sulfate proteoglycans, PG-I/biglycan and PG-II/decorin, by vascular smooth muscle and endothelial cells in culture

    J. Biol. Chem.

    (1991)
  • V. Kainulainen et al.

    Suppression of syndecan-1 expression in endothelial cells by tumor necrosis factor-α

    J. Biol. Chem.

    (1996)
  • K.J. Leco et al.

    Tissue inhibitor of metalloproteinase-3 (TIMP-3) is an extracellular matrix-associated protein with a distinctive pattern of expression in mouse cells and tissues

    J. Biol. Chem.

    (1994)
  • L. Mattila et al.

    Activation of tissue inhibitor of metalloproteinases-3 (TIMP-3) mRNA expression in scleroderma skin fibroblasts

    J. Invest. Dermatol.

    (1998)
  • L. Nelimarkka et al.

    Expression of small extracellular chondroitin/dermatan sulfate proteoglycans is differentially regulated in human endothelial cells

    J. Biol. Chem.

    (1997)
  • C.O. Savage et al.

    The role of the endothelium in systemic vasculitis

    J. Autoimmun.

    (1993)
  • M. Ståhle-Bäckdahl et al.

    Expression of 92-kDa type IV collagenase mRNA by eosinophils associated with basal cell carcinoma

    J. Invest. Dermatol.

    (1992)
  • W.G. Stetler-Stevenson et al.

    Tissue inhibitor of metalloproteinase (TIMP-2). A new member of the metalloproteinase inhibitor family

    J. Biol. Chem.

    (1989)
  • J. Westermarck et al.

    TNF-R55-specific form of human tumor necrosis factor-α induces collagenase gene expression by human skin fibroblasts

    J. Invest. Dermatol.

    (1995)
  • L. Badimon et al.

    Endothelium and atherosclerosis

    J. Hypertens.

    (1992)
  • D.J. Bidanset et al.

    Regulation of cell substrate adhesion: effects of small galactosaminoglycan-containing proteoglycans

    J. Cell Biol.

    (1992)
  • H. Birkedal-Hansen et al.

    Matrix metalloproteinases: a review

    Crit. Rev. Oral Biol. Med.

    (1993)
  • J.R. Bradley et al.

    Prolonged cytokine exposure causes a dynamic redistribution of endothelial cell adhesion molecules to intercellular junctions

    Lab. Invest.

    (1996)
  • F.M. Brennan et al.

    Reduction of serum matrix metalloproteinase 1 and matrix metalloproteinase 3 in rheumatoid arthritis patients following anti-tumour necrosis factor-α (cA2) therapy

    Br. J. Rheumatol.

    (1997)
  • D.F. Carmichael et al.

    Primary structure and cDNA cloning of human fibroblast collagenase inhibitor

  • C.J. Edgell et al.

    Permanent cell line expressing human factor VIII-related antigen established by hybridization

  • K. Elenius et al.

    Function of the syndecans — a family of cell surface proteoglycans

    J. Cell Sci.

    (1994)
  • H.D. Foda et al.

    Activation of human umbilical vein endothelial cell progelatinase A by phorbol myristate acetate: a protein kinase C-dependent mechanism involving a membrane-type matrix metalloproteinase

    Lab. Invest.

    (1996)
  • P. Fort et al.

    Various rat adult tissues express only one major mRNA species from the glyceraldehyde-3-phosphate-dehydrogenase multigenic family

    Nucleic Acids Res.

    (1985)
  • Z.S. Galis et al.

    Enhanced expression of vascular matrix metalloproteinases induced in vitro by cytokines and in regions of human atherosclerotic lesions

    Ann. NY Acad. Sci.

    (1995)
  • Z.S. Galis et al.

    Increased expression of matrix metalloproteinases and matrix degrading activity in vulnerable regions of human atherosclerotic plaques

    J. Clin. Invest.

    (1994)
  • Z.S. Galis et al.

    Microscopic localization of active proteases by in situ zymography: detection of matrix metalloproteinase activity in vascular tissue

    FASEB J.

    (1995)
  • I. Halpert et al.

    Matrilysin is expressed by lipid-laden macrophages at sites of potential rupture in atherosclerotic lesions and localizes to areas of versican deposition, a proteoglycan substrate for the enzyme

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