Inhibition of Vascular Smooth Muscle Cell Growth by Inhibition of Fibronectin Matrix Assembly

From the Department of Pathology, University of Chicago, Chicago, Ill.

Correspondence to Alex Morla, PhD, Department of Pathology, MC 6079, Committee on Cancer Biology, University of Chicago, 5841 S Maryland Ave, Chicago, IL 60637. E-mail amorla{at}midway.uchicago.edu

Abstract—The regulation of vascular smooth muscle cell (VSMC) proliferation by the fibronectin matrix was tested by treating human umbilical artery smooth muscle cells (HUASMCs) with a recombinant fragment of fibronectin (protein III₁-C) that has previously been shown to modulate fibronectin matrix assembly. III₁-C inhibited HUASMC proliferation by 75% to 90%. The inhibition of growth was time dependent; III₁-C had no effect on DNA synthesis after 0 to 5 hours of treatment but did have an effect at 24 hours and beyond. III₁-C did not stimulate apoptosis in these cells, indicating that the inhibition of proliferation was not due to an induction of programmed cell death. The effects of III₁-C on cell growth were only specific for normal diploid smooth muscle cells. III₁-C had no effect on the proliferation of IMR-90 fibroblasts, endothelial cells, NIH 3T3 cells, or the rat aortic smooth muscle cell line A7r5. However, III₁-C did inhibit proliferation by primary rat aortic smooth muscle cells. An analysis of HUASMC fibronectin receptor (integrin 5ß1) distribution revealed that III₁-C did not inhibit 5ß1 localization to focal contacts. Moreover, III₁-C had no effect on the relative expression levels of seven different integrin subunits on HUASMCs. However, III₁-C did inhibit fibronectin matrix assembly by rat aortic smooth muscle cells, HUASMCs, A7r5 cells, IMR-90 cells, and endothelial cells. An analysis of fibronectin synthesis indicated that the inhibition of fibronectin matrix assembly by III₁-C was not due solely to a decrease in fibronectin synthesis. Finally, treatment of HUASMCs with anti-fibronectin monoclonal antibody L8 (which is known to inhibit fibronectin matrix assembly) also decreased the rate of HUASMC DNA synthesis. These results demonstrate that III₁-C inhibits VSMC proliferation and suggest that this effect may be mediated by the inhibition of fibronectin matrix assembly.

Key Words: extracellular matrix • integrin • laminin • Matrigel

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