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Blood, Vol. 94 No. 8 (October 15), 1999:
pp. 2754-2766
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
Jordi Esparza,
Carme Vilardell,
Javier Calvo,
Manel Juan,
Jordi Vives,
Alvaro Urbano-Márquez,
Jordi Yagüe, and
Maria C. Cid
From the Departments of Internal Medicine and Immunology, Hospital
Clínic, University of Barcelona, IDIBAPS (Institut d'
Investigacions Biomèdiques August Pi i Sunyer), Barcelona, Spain;
and Immunology Unit, Hospital Germans Trias i Pujol, Badalona, Spain.
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  4, 5, and v
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.
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