The Migration of Human Smooth Muscle Cells In Vitro Is Mediated by Plasminogen Activation and Can Be Inhibited by α2-Macroglobulin Receptor Associated Protein

Monique J Wijnberg; Paul H A Quax; Nancy M E Nieuwenbroek; Jan H Verheijen

doi:10.1055/s-0038-1657646

Thrombosis and Haemostasis, Inhaltsverzeichnis

Thromb Haemost 1997; 78(02): 880-886
DOI: 10.1055/s-0038-1657646

Rapid Communication

Schattauer GmbH Stuttgart

The Migration of Human Smooth Muscle Cells In Vitro Is Mediated by Plasminogen Activation and Can Be Inhibited by α₂-Macroglobulin Receptor Associated Protein

Monique J Wijnberg

The Gaubius Laboratory, TNO-PG, Leiden, The Netherlands

,

Paul H A Quax

The Gaubius Laboratory, TNO-PG, Leiden, The Netherlands

,

Nancy M E Nieuwenbroek

The Gaubius Laboratory, TNO-PG, Leiden, The Netherlands

,

Jan H Verheijen

The Gaubius Laboratory, TNO-PG, Leiden, The Netherlands

› Institutsangaben

Abstract

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Summary

The plasminogen activation system is thought to be important in cell migration processes. A role for this system during smooth muscle cell migration after vascular injury has been suggested from several animal studies. However, not much is known about its involvement in human vascular remodelling. We studied the involvement of the plasminogen activation system in human smooth muscle cell migration in more detail using an in vitro wound assay and a matrix invasion assay. Inhibition of plasmin activity or inhibition of urokinase-type plasminogen activator (u-PA) activity resulted in approximately 40% reduction of migration after 24 h in the wound assay and an even stronger reduction (70-80%) in the matrix invasion assay. Migration of smooth muscle cells in the presence of inhibitory antibodies against tissue-type plasminogen activator (t-PA) was not significantly reduced after 24 h, but after 48 h a 30% reduction of migration was observed, whereas in the matrix invasion assay a 50% reduction in invasion was observed already after 24 h. Prevention of the interaction of u-PA with cell surface receptors by addition of soluble u-PA receptor or α₂-macroglobulin receptor associated protein (RAP) to the culture medium, resulted in a similar inhibition of migration and invasion. From these results it can be concluded that both u-PA and t-PA mediated plasminogen activation can contribute to in vitro human smooth muscle cell migration and invasion. Furthermore, the interaction between u-PA and its cell surface receptor appears also to be involved in this migration and invasion process. The inhibitory effects on migration and invasion by the addition of RAP suggests an involvement of a RAP sensitive receptor of the LDL receptor family, possibly the LDL-receptor related protein (LRP) and/or the VLDL receptor.

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References
1 Casscells W. Migration of smooth muscle and endothelial cells – critical events in restenosis. Circulation 1992; 86: 723-729
2 O’Brien ER, Schwartz SM. Update on the biology and clinical study of restenosis. Trends Cardiovasc Med 1994; 4: 169-178
3 Vassalli JD, Sappino AP, Belin D. The plasminogen activator/plasmin system. J Clin Invest 1991; 88: 1067-1072
4 Pepper MS, Belin D, Montesano R, Orci L, Vassalli JD. Transforming growth factor-beta-1 modulates basic fibroblast growth factor induced proteolytic and angiogenic properties of endothelial cells invitro. J Cell Biol 1990; 111: 743-755
5 Pepper MS, Sappino AP, Stocklin R, Montesano R, Orci L, Vassalli JD. Upregulation of urokinase receptor expression on migrating endothelial cells. J Cell Biol 1993; 122: 673-684
6 Clowes AW, Clowes MM, Au YPT, Reidy MA, Belin D. Smooth muscle cells express urokinase during mitogenesis and tissue-type plasminogen activator during migration in injured rat carotid artery. Circ Res 1990; 67: 61-67
7 Blasi F. Urokinase and urokinase receptor – a paracrine/autocrine system regulating cell migration and invasiveness. BioEssays 1993; 15: 105-111
8 Vassalli JD, Baccino D, Belin D. A cellular binding site for the Mr 55,000 form of the human plasminogen activator, urokinase. J Cell Biol 1985; 100: 86-92
9 Stoppelli MP, Corti A, Soffientini A, Cassani G, Blasi F, Associan RK. Differentiation enhanced binding of the amino terminal fragment of human urokinase-type plasminogen activator to a specific receptor on U937 monocytes. Proc Natl Acad Sci USA 1985; 82: 4939-4943
10 Cubellis MV, Wun TC, Blasi F. Receptor-mediated internalization and degradation of urokinase is caused by its specific inhibitor PAI-1. EMBO J 1990; 9: 1079-1085
11 Estreicher A, Muhlhauser J, Carpentier JL, Orci L, Vassalli JD. The receptor for urokinase type plasminogen activator polarizes expression of the protease to the leading edge of migrating monocytes and promotes degradation of enzyme inhibitor complexes. J Cell Biol 1990; 111: 783-792
12 Blasi F, Conese M, Moller LB, Pedersen N, Cavallaro U, Cubellis MV, Fazioli F, Hemandezmarrero L, Limongi P, Munozcanoves P, Resnati M, Riittinen L, Sidenius N, Soravia E, Soria MR, Stoppelli MP, Talarico D, Teesalu T, Valcamonica S. The urokinase receptor: structure, regulation and inhibitor-mediated internalization. Fibrinolysis 1994; 08 (Suppl. 01) 182-188
13 Danø K, Behrendt N, Brunner N, Ellis V, Ploug M, Pyke C. The urokinase receptor. Protein structure and role in plasminogen activation and cancer invasion. Fibrinolysis 1994; 08 (Suppl. 01) 189-203
14 Vassalli JD. The urokinase receptor. Fibrinolysis 1994; 08 (Suppl. 01) 172-181
15 Nykjaer A, Petersen CM, Moller B, Jensen PH, Moestrup SK, Holtet TL, Etzerodt M, Thogersen HC, Munch M, Andreasen PA, Glieman J. Purified a₂-Macroglobulin Receptor/LDL receptor-related protein binds urokinase activator inhibitor type-1 complex – evidence that the a₂-Macroglobulin receptor mediates cellular degradation of urokinase receptor-bound complexes. J Biol Chem 1992; 267: 14543-14546
16 Heegaard CW, Wiborg SimonsenAC, Oka K, Kjøller L, Christensen A, Madsen B, Ellgaard L, Chan L, Andreasen PA. Very low density lipoprotein receptor binds and mediates endocytosis of urokinase-type plasminogen activator-type-1 plasminogen activator inhibitor complex. J Biol Chem 1995; 270: 20855-20861
17 Strickland DK, Kounnas MZ, Williams SE, Argraves WS. LDL receptor-related protein (LRP): a multiligand receptor. Fibrinolysis 1994; 08 (Suppl. 01) 204-215
18 Moestrup SK. The α₂-macroglobulin receptor and epithelial glycoprotein-330: two giant receptors mediating endocytosis of multiple ligands. Bio-chim Biophys Acta 1994; 1197: 197-213
19 Conese M, Nykjaer A, Petersen CM, Cremona O, Pardi R, Andreasen PA, Gliemann J, Christensen El, Blasi F. Alpha-2 Macroglobulin receptor/Ldl receptor-related protein (Lrp)-dependent internalization of the urokinase receptor. J Cell Biol 1995; 131: 1609-1622
20 Grobmyer SR, Kuo A, Orishimo M, Okada SS, Cines DB, Bamathan ES. Determinants of binding and internalization of tissue-type plasminogen activator by human vascular smooth muscle and endothelial cells. J Biol Chem 1993; 268: 13291-13300
21 Ossowski L, Clunie G, Masucci MT, Blasi F. In vivo paracrine interaction between urokinase and its receptor: effect on tumor cell invasion. J Cell Biol 1991; 115: 1107-1112
22 Quax PHA, Pedersen N, Masucci MT, Weening-Verhoeff EJD, Danø K, Verheijen JH, Blasi F. Complementation between urokinase-producing and receptor producing cells in extracellular matrix degradation. Cell Regul 1991; 2: 793-803
23 Quax PHA, Boxman ILA, Van KesterenCAM, Verheijen JH, Ponec M. Plasminogen activators are involved in keratinocyte and fibroblast migration in wounded cultures in vitro. Fibrinolysis 1994; 8: 221-228
24 Koolwijk P, Vanerck MGM, Devree WJA, Vermeer MA, Weich HA, Hanemaaijer R, Vanhinsbergh VWM. Cooperative effect of TNF alpha, bFGF, and VEGF on the formation of tubular structures of human micro-vascular endothelial cells in a fibrin matrix. Role of urokinase activity. J Cell Biol 1996; 122: 673-684
25 Schleef RR, Birdwell CR. The effects of proteases on endothelial cell migration in vitro. Exp Cell Res 1982; 141: 503-508
26 Jackson CL, Raines EW, Ross R, Reidy MA. Role of endogenous platelet-derived growth factor in arterial smooth muscle cell migration after balloon catheter injury. Arterioscl Thromb 1993; 13: 1218-1226
27 Carmeliet P, Collen D. Evaluation of the plasminogen/plasmin system in transgenic mice. Fibrinolysis 1994; 08 (Suppl. 01) 269-276
28 Carmeliet P, Moons L, Dewerchin M, Ploplis V, De MolM, Stassen JM, Declerq C, Plow E, Gerard R, Collen D. Receptor-independent plasminogen activation by urokinase-type plasminogen activator promotes, and adenoviral PAI-1 gene transfer inhibits arterial neointima formation in mice. Fibrinolysis 1996; 10 (Suppl. 03) 19
29 Noda-Heiny H, Daugherty A, Sobel BE. Augmented urokinase receptor expression in atheroma. Arterioscler Thromb Vase Biol 1995; 15: 37-43
30 Lupu F, Heim D, Bachmann F, Kruithof EKO. Expression of LDL receptor-related protein/α₂-macroglobulin receptor in human normal and atherosclerotic arteries. Arteriocler Thromb 1994; 14: 1438-1444
31 Lupu F, Heim DA, Bachmann F, Humi M, Kakkar VV, Kruithof EKO. Plasminogen activator expression in human atherosclerotic lesions. Arterioscler Thromb Vase Biol 1995; 15: 1444-1455
32 Raghunath PN, Tomaszewski JE, Brady ST, Caron RJ, Okada SS, Bamathan ES. Plasminogen activator system in human coronary atherosclerosis. Arterioscler Thromb Vase Biol 1995; 15: 1432-1443
33 Padró T, Emeis JJ, Steins M, Schmid KW, Kienast J. Quantification of plasminogen activators and their inhibitors in the aortic vessel wall in relation to the presence and severity of atherosclerotic disease. Arterioscler Thromb Vase Biol 1995; 15: 893-902
34 Wijnberg MJ, Nieuwenbroek NME, Slomp J, Quax PHA, Verheijen JH. Urokinase and tissue-type plasminogen activator stimulate human vascular smooth muscle cell migration. Fibrinolysis 1996; 10 (Suppl. 02) 75-78
35 Jaffe EA, Nachmann RL, Becker CG. Culture of endothelial cells derived from umbilical veins – Identification by morphologic and immunologic criteria. J Clin Invest 1973; 52: 2745-2756
36 Kocan RM, Moss NS, Benditt EP. Human arterial wall cells and tissues in culture. Methods in Cell Biology 1980; 21A: 153-166
37 Rijken DC, Wijngaards G, Welbergen J. Immunological characterization of plasminogen activator activities in human tissues and body fluids. J Lab Clin Med 1981; 97: 477-486
38 Binnema DJ, Van IerselJJL, Dooijewaard G. Quantitation of urokinase antigen in plasma and culture media by use of an ELISA. Thromb Res 1986; 43: 569-577
39 Chucholowski N, Schmitt M, Rettenberger D, Schuren E, Moniwa N, Goretzki L, Wilhelm O, Weidle U, Janicke F, Graeff H. Flow cyto-fluorometric analysis of the urokinase receptor (uPAR) on tumor cells by fluorescent uPA-ligand or monoclonal antibody 3936. Fibrinolysis 1992; 06 (Suppl. 04) 95-102
40 Herz J, Hamann U, Rogne S, Myklebost O, Gausepohl H, Stanley KK. Surface location and high affinity for calcium of a 500-kd liver membrane protein closely related to the LDL-receptor suggest a physiological role as lipoprotein receptor. EMBO J 1988; 7: 4119-4127
41 Denizot F, Lang R. Rapid colorimetric assay for cell growth and survival. Modifications to the tetrazolium dye procedure giving improved sensitivity and reliability. J Immunol Methods 1986; 89: 271-277
42 Laemmli UK. Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature 1970; 227: 680-685
43 Granelli-Pipemo A, Reich E. A study of proteases and protease inhibitor complexes in biological fluids. J Exp Med 1978; 148: 223-234
44 Sperti G, Van LeeuwenRTJ, Quax PHA, Maseri A, Kluft C. Cultured rat aortic vascular smooth muscle cells digest naturally produced extracellular matrix. Involvement of plasminogen-dependent and plasminogen-independent pathways. Circ Res 1992; 71: 385-392
45 Pauly RR, Passaniti A, Bilato C, Monticone R, Cheng L, Papadopoulos N, Gluzland YA, Smith L, Weinstein C, Lakatta EG, Crow MT. Migration of cultured vascular smooth muscle cells through a basement membrane requires type IV collagenase activity and is inhibited by cellular differentiation. Circ Res 1994; 75: 41-54
46 Bendeck M, Zempo N, Clowes MM, Galardy R, Reidy MA. Smooth muscle cell migration and matrix metalloproteinase expression after arterial injury in the rat. Circ Res 1994; 75: 539-545
47 Okada SS, Tomaszewski JE, Bamathan ES. Migrating vascular smooth muscle cells polarize cell surface urokinase receptors after injury in vitro. Exp Cell Res 1995; 217: 180-187
48 Wei Y, Waltz DA, Rao N, Drummond RJ, Rosenberg J, Chapman HA. Identification of the urokinase receptor as an adhesion receptor for vitronectin. J Biol Chem 1994; 269: 32380-32388