Fibronectin upregulates gelatinase B (MMP-9) and induces coordinated expression of gelatinase A (MMP-2) and its activator MT1-MMP (MMP-14) by human T lymphocyte cell lines. A process repressed through RAS/MAP kinase signaling pathways

Blood. 1999 Oct 15;94(8):2754-66.

Abstract

T-lymphocyte migration into tissues requires focal degradation of the basement membrane. In this study, we show that transient adherence to fibronectin induces the production of activated forms of matrix metalloproteinase-2 (MMP-2) and MMP-9, as well as downregulation of tissue inhibitor of metalloproteinase-2 (TIMP-2) by T-cell lines. MMP-2 activation was likely achieved by inducing a coordinated expression of membrane-type matrix metalloproteinase-1 (MMP-14), a major activator of MMP-2. Blocking monoclonal antibodies against alpha4, alpha5, and alphav integrins strongly reduced MMP-2 and MMP-9 production induced by fibronectin. Disrupting actin cytoskeleton organization by cytochalasin D strongly enhanced fibronectin-induced MMP-2 and MMP-9 expression. Inhibiting Src tyrosine kinases with herbimycin A reduced MMP-2 and MMP-9 production with no effect on cell attachment. By contrast, G-protein inhibition by pertussis toxin, or transfection with a dominant negative mutant of Ha-Ras strongly increased fibronectin-induced MMP-2 and MMP-9. Inhibition of PI3 kinase, MAPkinase (MEK1), or p38 MAPkinase by wortmannin, PD 98059, or SB 202190, respectively, strongly promoted fibronectin-induced MMP2 and MMP-9. Cells at high density lost their ability to synthesize MMP-2 and MMP-9 in response to fibronectin and MMP expression was restored by transfection with a dominant-negative mutant of Ha-Ras or by treatment with wortmannin, PD 98059, or SB 202190. Our findings suggest that adhesion to fibronectin transduces both stimulatory (through Src-type tyrosin kinases) and inhibitory signals (through Ras/MAPKinase signaling pathways) for MMP-2 and MMP-9 expression by T lymphocytes and that their relative predominance is regulated by additional stimuli related to cell adhesion, motility, and growth.

Publication types

  • Comparative Study
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Androstadienes / pharmacology
  • Antibodies, Monoclonal / pharmacology
  • Benzoquinones
  • Cell Adhesion / drug effects
  • Enzyme Activation / drug effects
  • Enzyme Inhibitors / pharmacology
  • Fibronectins / pharmacology*
  • Flavonoids / pharmacology
  • GTP-Binding Proteins / antagonists & inhibitors
  • Gene Expression Regulation / drug effects*
  • Gene Expression Regulation, Neoplastic / drug effects
  • Genes, ras
  • Humans
  • Imidazoles / pharmacology
  • Integrins / antagonists & inhibitors
  • Integrins / immunology
  • Jurkat Cells / drug effects
  • Jurkat Cells / metabolism
  • Lactams, Macrocyclic
  • MAP Kinase Signaling System*
  • Matrix Metalloproteinase 2 / biosynthesis*
  • Matrix Metalloproteinase 2 / genetics
  • Matrix Metalloproteinase Inhibitors*
  • Matrix Metalloproteinases / biosynthesis*
  • Matrix Metalloproteinases / genetics
  • Matrix Metalloproteinases, Membrane-Associated
  • Metalloendopeptidases*
  • Neoplasm Proteins / biosynthesis
  • Neoplasm Proteins / genetics
  • Neoplasm Proteins / physiology
  • Peptide Fragments / pharmacology
  • Pertussis Toxin
  • Phosphoinositide-3 Kinase Inhibitors
  • Protein Structure, Tertiary
  • Proto-Oncogene Proteins p21(ras) / antagonists & inhibitors
  • Proto-Oncogene Proteins p21(ras) / physiology*
  • Pyridines / pharmacology
  • Quinones / pharmacology
  • Rifabutin / analogs & derivatives
  • T-Lymphocytes / drug effects*
  • T-Lymphocytes / metabolism
  • Tissue Inhibitor of Metalloproteinase-1 / biosynthesis
  • Tissue Inhibitor of Metalloproteinase-1 / genetics
  • Tissue Inhibitor of Metalloproteinase-2 / biosynthesis
  • Tissue Inhibitor of Metalloproteinase-2 / genetics
  • Transfection
  • Tumor Cells, Cultured
  • Virulence Factors, Bordetella / pharmacology
  • Wortmannin
  • src-Family Kinases / antagonists & inhibitors
  • src-Family Kinases / physiology

Substances

  • Androstadienes
  • Antibodies, Monoclonal
  • Benzoquinones
  • Enzyme Inhibitors
  • Fibronectins
  • Flavonoids
  • Imidazoles
  • Integrins
  • Lactams, Macrocyclic
  • Matrix Metalloproteinase Inhibitors
  • Neoplasm Proteins
  • Peptide Fragments
  • Phosphoinositide-3 Kinase Inhibitors
  • Pyridines
  • Quinones
  • Tissue Inhibitor of Metalloproteinase-1
  • Virulence Factors, Bordetella
  • Tissue Inhibitor of Metalloproteinase-2
  • Rifabutin
  • herbimycin
  • Pertussis Toxin
  • src-Family Kinases
  • Matrix Metalloproteinases
  • Matrix Metalloproteinases, Membrane-Associated
  • Metalloendopeptidases
  • Matrix Metalloproteinase 2
  • GTP-Binding Proteins
  • HRAS protein, human
  • Proto-Oncogene Proteins p21(ras)
  • 4-(4-fluorophenyl)-2-(4-hydroxyphenyl)-5-(4-pyridyl)imidazole
  • 2-(2-amino-3-methoxyphenyl)-4H-1-benzopyran-4-one
  • Wortmannin