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

This article has been cited by other articles:

				P. Fogelstrand, C. C. Feral, R. Zargham, and M. H. Ginsberg Dependence of proliferative vascular smooth muscle cells on CD98hc (4F2hc, SLC3A2) J. Exp. Med., October 26, 2009; 206(11): 2397 - 2406. [Abstract] [Full Text] [PDF]

				H.-Y. Chiang, V. A. Korshunov, A. Serour, F. Shi, and J. Sottile Fibronectin Is an Important Regulator of Flow-Induced Vascular Remodeling Arterioscler Thromb Vasc Biol, July 1, 2009; 29(7): 1074 - 1079. [Abstract] [Full Text] [PDF]

				K. Khavandi, A. S. Greenstein, K. Sonoyama, S. Withers, A. Price, R. A. Malik, and A. M. Heagerty Myogenic tone and small artery remodelling: insight into diabetic nephropathy Nephrol. Dial. Transplant., February 1, 2009; 24(2): 361 - 369. [Full Text] [PDF]

				C.-H. Chen, H.-J. Wei, W.-W. Lin, I. Chiu, S.-M. Hwang, C.-C. Wang, W.-Y. Lee, Y. Chang, and H.-W. Sung Porous tissue grafts sandwiched with multilayered mesenchymal stromal cell sheets induce tissue regeneration for cardiac repair Cardiovasc Res, October 1, 2008; 80(1): 88 - 95. [Abstract] [Full Text] [PDF]

				X. Zhou, R. G. Rowe, N. Hiraoka, J. P. George, D. Wirtz, D. F. Mosher, I. Virtanen, M. A. Chernousov, and S. J. Weiss Fibronectin fibrillogenesis regulates three-dimensional neovessel formation Genes & Dev., May 1, 2008; 22(9): 1231 - 1243. [Abstract] [Full Text] [PDF]

				K. Sonoyama, A. Greenstein, A. Price, K. Khavandi, and T. Heagerty Review: Vascular remodeling: implications for small artery function and target organ damage Therapeutic Advances in Cardiovascular Disease, December 1, 2007; 1(2): 129 - 137. [Abstract] [PDF]

				J. Sottile, F. Shi, I. Rublyevska, H.-Y. Chiang, J. Lust, and J. Chandler Fibronectin-dependent collagen I deposition modulates the cell response to fibronectin Am J Physiol Cell Physiol, December 1, 2007; 293(6): C1934 - C1946. [Abstract] [Full Text] [PDF]

				H. Meng, Z. Wang, Y. Hoi, L. Gao, E. Metaxa, D. D. Swartz, and J. Kolega Complex Hemodynamics at the Apex of an Arterial Bifurcation Induces Vascular Remodeling Resembling Cerebral Aneurysm Initiation Stroke, June 1, 2007; 38(6): 1924 - 1931. [Abstract] [Full Text] [PDF]

				L. Gui, K. Wojciechowski, C. D. Gildner, H. Nedelkovska, and D. C. Hocking Identification of the Heparin-binding Determinants within Fibronectin Repeat III1: ROLE IN CELL SPREADING AND GROWTH J. Biol. Chem., November 17, 2006; 281(46): 34816 - 34825. [Abstract] [Full Text] [PDF]

				P. Brassard, F. Amiri, G. Thibault, and E. L. Schiffrin Role of Angiotensin Type-1 and Angiotensin Type-2 Receptors in the Expression of Vascular Integrins in Angiotensin II-Infused Rats Hypertension, January 1, 2006; 47(1): 122 - 127. [Abstract] [Full Text] [PDF]

				M. Otis, S. Campbell, M. D. Payet, and N. Gallo-Payet Angiotensin II Stimulates Protein Synthesis and Inhibits Proliferation in Primary Cultures of Rat Adrenal Glomerulosa Cells Endocrinology, February 1, 2005; 146(2): 633 - 642. [Abstract] [Full Text] [PDF]

				A. Ambesi, R. M. Klein, K. M. Pumiglia, and P. J. McKeown-Longo Anastellin, a Fragment of the First Type III Repeat of Fibronectin, Inhibits Extracellular Signal-Regulated Kinase and Causes G1 Arrest in Human Microvessel Endothelial Cells Cancer Res., January 1, 2005; 65(1): 148 - 156. [Abstract] [Full Text] [PDF]

				R. M. Klein, M. Zheng, A. Ambesi, L. Van De Water, and P. J. McKeown-Longo Stimulation of extracellular matrix remodeling by the first type III repeat in fibronectin J. Cell Sci., November 15, 2003; 116(22): 4663 - 4674. [Abstract] [Full Text] [PDF]

				S. H. Wilson, A. V. Ljubimov, A. O. Morla, S. Caballero, L. C. Shaw, P. E. Spoerri, R. W. Tarnuzzer, and M. B. Grant Fibronectin Fragments Promote Human Retinal Endothelial Cell Adhesion and Proliferation and ERK Activation through {alpha}5{beta}1 Integrin and PI 3-Kinase Invest. Ophthalmol. Vis. Sci., April 1, 2003; 44(4): 1704 - 1715. [Abstract] [Full Text] [PDF]

				J. Sottile and D. C. Hocking Fibronectin Polymerization Regulates the Composition and Stability of Extracellular Matrix Fibrils and Cell-Matrix Adhesions Mol. Biol. Cell, October 1, 2002; 13(10): 3546 - 3559. [Abstract] [Full Text] [PDF]

				D. Simper, P. G. Stalboerger, C. J. Panetta, S. Wang, and N. M. Caplice Smooth Muscle Progenitor Cells in Human Blood Circulation, September 3, 2002; 106(10): 1199 - 1204. [Abstract] [Full Text] [PDF]

				G. P. Rossi, M. Cavallin, A. S Belloni, G. Mazzocchi, G. G Nussdorfer, A. C Pessina, and S. Sartore Aortic smooth muscle cell phenotypic modulation and fibrillar collagen deposition in angiotensin II-dependent hypertension Cardiovasc Res, July 1, 2002; 55(1): 178 - 189. [Abstract] [Full Text] [PDF]

				I. Wierzbicka-Patynowski and J. E. Schwarzbauer Regulatory Role for Src and Phosphatidylinositol 3-Kinase in Initiation of Fibronectin Matrix Assembly J. Biol. Chem., May 24, 2002; 277(22): 19703 - 19708. [Abstract] [Full Text] [PDF]

				E. P. Moiseeva Adhesion receptors of vascular smooth muscle cells and their functions Cardiovasc Res, December 1, 2001; 52(3): 372 - 386. [Abstract] [Full Text] [PDF]

				J. W. Fischer, M. G. Kinsella, B. Levkau, A. W. Clowes, and T. N. Wight Retroviral Overexpression of Decorin Differentially Affects the Response of Arterial Smooth Muscle Cells to Growth Factors Arterioscler Thromb Vasc Biol, May 1, 2001; 21(5): 777 - 784. [Abstract] [Full Text] [PDF]

				Y. Wen, J. Gu, Y. Liu, P. H. Wang, Y. Sun, and J. L. Nadler Overexpression of 12-Lipoxygenase Causes Cardiac Fibroblast Cell Growth Circ. Res., January 19, 2001; 88(1): 70 - 76. [Abstract] [Full Text] [PDF]

				H. D. Intengan and E. L. Schiffrin Structure and Mechanical Properties of Resistance Arteries in Hypertension : Role of Adhesion Molecules and Extracellular Matrix Determinants Hypertension, September 1, 2000; 36(3): 312 - 318. [Abstract] [Full Text] [PDF]

				M. G. Kinsella, J. W. Fischer, D. P. Mason, and T. N. Wight Retrovirally Mediated Expression of Decorin by Macrovascular Endothelial Cells. EFFECTS ON CELLULAR MIGRATION AND FIBRONECTIN FIBRILLOGENESIS IN VITRO J. Biol. Chem., April 28, 2000; 275(18): 13924 - 13932. [Abstract] [Full Text] [PDF]

				D. C. Hocking, J. Sottile, and K. J. Langenbach Stimulation of Integrin-mediated Cell Contractility by Fibronectin Polymerization J. Biol. Chem., March 31, 2000; 275(14): 10673 - 10682. [Abstract] [Full Text] [PDF]

				R. K. Suniara, E. J. Jenkinson, and J. J.T. Owen An Essential Role for Thymic Mesenchyme in Early T Cell Development J. Exp. Med., March 20, 2000; 191(6): 1051 - 1056. [Abstract] [Full Text] [PDF]

				J. G. Pickering, L. H. Chow, S. Li, K. A. Rogers, E. F. Rocnik, R. Zhong, and B. M. C. Chan {alpha}5{beta}1 Integrin Expression and Luminal Edge Fibronectin Matrix Assembly by Smooth Muscle Cells after Arterial Injury Am. J. Pathol., February 1, 2000; 156(2): 453 - 465. [Abstract] [Full Text] [PDF]

				J Sottile, D. Hocking, and K. Langenbach Fibronectin polymerization stimulates cell growth by RGD-dependent and -independent mechanisms J. Cell Sci., January 12, 2000; 113(23): 4287 - 4299. [Abstract] [PDF]

				K. J. Langenbach and J. Sottile Identification of Protein-disulfide Isomerase Activity in Fibronectin J. Biol. Chem., March 12, 1999; 274(11): 7032 - 7038. [Abstract] [Full Text] [PDF]

				R. Ross Atherosclerosis -- An Inflammatory Disease N. Engl. J. Med., January 14, 1999; 340(2): 115 - 126. [Full Text] [PDF]

				R. Christopher, S. Judge, P. Vincent, P. Higgins, and P. McKeown-Longo The amino-terminal matrix assembly domain of fibronectin stabilizes cell shape and prevents cell cycle progression J. Cell Sci., January 10, 1999; 112(19): 3225 - 3235. [Abstract] [PDF]

				S. Bourdoulous, G. Orend, D. A. MacKenna, R. Pasqualini, and E. Ruoslahti Fibronectin Matrix Regulates Activation of RHO and CDC42 GTPases and Cell Cycle Progression J. Cell Biol., October 5, 1998; 143(1): 267 - 276. [Abstract] [Full Text] [PDF]

				J. L. Sechler and J. E. Schwarzbauer Control of Cell Cycle Progression by Fibronectin Matrix Architecture J. Biol. Chem., October 2, 1998; 273(40): 25533 - 25536. [Abstract] [Full Text] [PDF]

				J Sottile, D. Hocking, and P. Swiatek Fibronectin matrix assembly enhances adhesion-dependent cell growth J. Cell Sci., January 10, 1998; 111(19): 2933 - 2943. [Abstract] [PDF]

				K. Tamura, Y. E. Chen, M. Lopez-Ilasaca, L. Daviet, N. Tamura, T. Ishigami, M. Akishita, I. Takasaki, Y. Tokita, R. E. Pratt, et al. Molecular Mechanism of Fibronectin Gene Activation by Cyclic Stretch in Vascular Smooth Muscle Cells J. Biol. Chem., October 27, 2000; 275(44): 34619 - 34627. [Abstract] [Full Text] [PDF]

				B. R. Tomasini-Johansson, N. R. Kaufman, M. G. Ensenberger, V. Ozeri, E. Hanski, and D. F. Mosher A 49-Residue Peptide from Adhesin F1 of Streptococcus pyogenes Inhibits Fibronectin Matrix Assembly J. Biol. Chem., June 22, 2001; 276(26): 23430 - 23439. [Abstract] [Full Text] [PDF]