Abstract
The plasminogen activation (PA) system plays an important role in tumor invasion by initiating pericellular proteolysis of the extracellular matrix (ECM) and inducing cell migration. Malignant brain tumors overexpress PA members and characteristically invade by migrating on ECM-producing white matter tracts and blood vessel walls. To determine whether urokinase-type plasminogen activator (uPA) and its receptor (uPAR) directly modulate the migration of brain tumor cells, we examined six human brain tumor cell lines, 2 astrocytomas (SW1088, SW1783), 2 medullobastomas (Daoy, D341Med), and 2 glioblastomas (U87MG, U118MG), for their surface uPAR expression, endogenous PA activity, and functional proteolytic activity by an ECM-degradation assay. Migration on Transwell membranes and invasion of Matrigel was then tested by pre-incubating the cells with increasing concentrations of either uPA, the proteolytically inactive amino-terminal fragment (ATF) of uPA, or the uPAR cleaving enzyme, phosphatidylinositol-specific phospholipase C (PI-PLC).
All of the cell lines, except D341Med, express surface uPAR protein and uPA activity. High levels of uPAR and uPA activity correlated with cellular degradation of ECM, cell migration, and Matrigel invasion. Cell migration and invasion were enhanced by uPA or ATF in a dose dependent manner, while PI-PLC treatment abolished the uPA effect and inhibited migration and invasion. We conclude that ligation of uPAR by uPA directly induces brain tumor cell migration, independent of uPA-mediated proteolysis; and in concert with ECM degradation, markedly enhances invasion. Conversely, removing membrane bound uPAR from the surface of the cells studied inhibited their ability to migrate and invade even in the presence of proteolytically active uPA.
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Quax PH, van Muijen GN, Weening Verhoeff EJ, Lund LR, Dano K, Ruiter DJ, Verheijen JH: Metastatic behavior of human melanoma cell lines in nude mice correlates with urokinase-type plasminogen activator, its type-1 inhibitor, and urokinase-mediated matrix degradation. J Cell Biol 115: 191–199, 1991
Mignatti P, Robbins E, Rifkin DB: Tumor invasion through the human amniotic membrane: requirement for a proteinase cascade. Cell 47: 487–498, 1986
Dano K, Andreasen PA, Grondahl Hansen J, Kristensen P, Nielsen LS, Skriver L: Plasminogen activators, tissue degradation, and cancer. Adv Cancer Res 44: 139–266, 1985
Costantini V, Sidoni A, Deveglia R, Cazzato OA, Bellezza G, Ferri I, Bucciarelli E, Nenci GG: Combined overexpression of urokinase, urokinase receptor, and plasminogen activator inhibitor-1 is associated with breast cancer progression: an immunohistochemical comparison of normal, benign, and malignant breast tissues. Cancer 77: 1079–1088, 1996
Pedersen H, Brunner N, Francis D, Osterlind K, Ronne E, Hansen HH, Dano K, Grondahl Hansen J: Prognostic impact of urokinase, urokinase receptor, and type 1 plasminogen activator inhibitor in squamous and large cell lung cancer tissue. Cancer Res 54: 4671–4675, 1994
Kohga S, Harvey SR, Weaver RM, Markus G: Localization of plasminogen activators in human colon cancer by immunoperoxidase staining. Cancer Res 45: 1787–1796, 1985
Blasi F, Verde P: Urokinase-dependent cell surface proteolysis and cancer. Semin Cancer Biol 1: 117–126, 1990
Polleanen J, Stephens RW, Vaheri A: Directed plasminogen activation at the surface of normal and malignant cells. Adv Cancer Res 57: 273–328, 1991
DeClerck YA, Laug WE: The role of the extracellular matrix in tumor invasion, metastasis and angiogenesis. In: Teicher BA (ed) Drug Resistance in Oncology. Marcel Dekker, New York 121–163, 1993
Mazzieri R, Masiero L, Zanetta L, Monea S, Onisto M, Garbisa S, Mignatti P: Control of type IV collagenase activity by components of the urokinase-plasmin system: a regulatory mechanism with cell-bound reactants. EMBO J 16: 2319–2332, 1997
Wei Y, Waltz DA, Rao N, Drummond RJ, Rosenberg S, Chapman HA: Identification of the urokinase receptor as an adhesion receptor for vitronectin. J Biol Chem 269: 32380–32388, 1994
Wei Y, Lukashev M, Simon DI, Bodary SC, Rosenberg S, Doyle MV, Chapman HA: Regulation of integrin function by the urokinase receptor. Science 273: 1551–1555, 1996
Nakano A, Tani E, Miyazaki K, Yamamoto Y, Furuyama J: Matrix metalloproteinases and tissue inhibitors of metalloproteinases in human gliomas. J Neurosurg 83: 298–307, 1995
Yamamoto M, Sawaya R, Mohanam S, Bindal AK, Bruner JM, Oka K, Rao VH, Tomonaga M, Nicholson GL, Rao JS: Expression and localization of urokinase-type plasminogen activator in human astrocytomas in vivo. Cancer Res 54: 3656–3661, 1994
Gladson CL, Pijuan Thompson V, Olman MA, Gillespie GY, Yacoub IZ: Up-regulation of urokinase and urokinase receptor genes in malignant astrocytoma. Am J Pathol 146: 1150–1160, 1995
Pollack IF: Brain tumors in children. N Engl J Med 331: 1500–1507, 1994
Johnson JH Jr, Phillips PC: Malignant gliomas in children. Cancer Invest 14: 609–621, 1996
Mohanam S, Sawaya R, McCutcheon I, Ali Osman F, Boyd D, Rao JS: Modulation of in vitro invasion of human glioblastoma cells by urokinase-type plasminogen activator receptor antibody. Cancer Res 53: 4143–4147, 1993
Paulus W, Bauer I, Beutler AS, Reeves SA: Diffuse brain invasion of glioma cells requires beta 1 integrins. Lab Invest 75: 819–826, 1996
Laug WE, Cao XR, Yu YB, Shimada H, Kruithof EK: Inhibition of invasion of HT1080 sarcoma cells expressing recombinant plasminogen activator inhibitor 2. Cancer Res 53: 6051–6057, 1993
Laug WE, Wang K, Mundi R, Rideout W, Kruithof EK, Bogenmann E: Clonal variation of expression of the genes coding for plasminogen activators, their inhibitors and the urokinase receptor in HT1080 sarcoma cells. Int J Cancer 52: 298–304, 1992
Laug WE, Weinblatt ME, Jones P: Endothelial cells degrade extracellular matrix proteins produced in vitro. Thromb Haemost 54: 498–502, 1985
Jones PA, DeClerck Y: Destruction of extracellular matrices containing glycoproteins, clastin, and collagen by metastatic human tumor cells. Cancer Res 40: 3222–3227, 1980
Stahl A, Mueller BM: Binding of urokinase to its receptor promotes migration and invasion of human melanoma cells in vitro. Cancer Res 54: 3066–3067, 1994
Repesh L: A new in vitro assay for quantitating tumor cell invasion. Invasion Metastasis 9: 192–208, 1989
Shirasuna K, Saka M, Hayashido Y, Yoshioka H, Sugiura T, Matsuya T: Extracellular matrix production and degradation by adenoid cystic carcinoma cells: participation of plasminogen activator and its inhibitor in matrix degradation. Cancer Res 53: 147–152, 1993
Gudewicz PW, Gilboa N: Human urokinase-type plasminogen activator stimulates chemotaxis of human neutrophils. Biochem Biophys Res Commun 147: 1176–1181, 1987
Busso N, Masur SK, Lazega D, Waxman S, Ossowski L: Induction of cell migration by pro-urokinase binding to its receptor: possible mechanism for signal transduction in human epithelial cells. J Cell Biol 126: 259–270, 1994
Kook YH, Adamski J, Zelent A, Ossowski: The effect of anti-sense inhibition of urokinase receptor in human squamous cell carcinoma on malignancy. EMBO J 13: 3983–3991, 1994
Kariko K, Kuo A, Boyd D, Okada SS, Cines DB, Barnathan ES: Overexpression of urokinase receptor increases matrix invasion without altering cell migration in a human oseosarcoma cell line. Cancer Res 53: 3109–3117, 1993
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 100: 86–92, 1985
Ploug M, Behrendt N, Lober D, Dano K: Protein structure and membrane anchorage of the cellular receptor for urokinase-type plasminogen activator. Semin Thromb Hemost 17: 183–193, 1991
Crowley CW, Cohen RL, Lucas BK, Liu G, Shuman MA, Levinson AD: Prevention of metastasis by inhibition of the urokinase receptor. Proc Natl Acad Sci USA 90: 5021–5025, 1993
Odekon LE, Sato Y, Rifkin DB: Urokinase-type plasminogen mediates basic fibroblast growth factor-induced bovine endothelial cell migration independent of its proteolytic activity. J Cell Physiol 150: 258–263, 1992
Waltz DA, Chapman HA: Reversible cellular adhesion to vitronectin linked to urokinase receptor occupancy. J Biol Chem 269: 14746–14750, 1994
Bohuslav J, Horejsi V, Hansmann C, Stockl J, Weidle UH, Majdic O, Bartke I, Knapp W, Stockinger H: Urokinase plasminogen activator receptor, beta 2-integrins, and Src-kinases within a single receptor complex of human monocytes. J Exp Med 181: 1381–1390, 1995
Giese A, Rief MD, Loo MA, Berens ME: Determinants of human astrocytoma migration. Cancer Res 54: 3897–3904, 1994
Paulus W, Tonn JC: Basement membrane invasion of glioma cells mediated by integrin receptors. J Neurosurg 80: 515–519, 1994
Appella E, Robinson EA, Ullrich SJ, Stoppelli MP, Corti A, Cassani G, Blasi F: The receptor-binding sequence of urokinase. A biological function for the growth-factor module of proteases. J Biol Chem 262: 4437–4440, 1987
Del Rosso M, Fibbi G, Dini G, Grappone C, Pucci M, Caldini R, Magnelli L, Fimiani M, Lotti T, Panconesi E: Role of specific membrane receptors in urokinase-dependent migration of human Keratinocytes. J Invest Dermatol 94: 310–316, 1990
Mohanam S, Chintala SK, Go Y, Bhattacharya A, Venkaiah B, Boyd D, Gokaslan ZL, Sawaya R, Rao JS: In vitro inhibition of human glioblastoma cell line invasiveness by antisense uPA receptor. Oncogene 14: 1351–1359, 1997
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MacDonald, T.J., DeClerck, Y.A. & Laug, W.E. Urokinase induces receptor mediated brain tumor cell migration and invasion. J Neurooncol 40, 215–226 (1998). https://doi.org/10.1023/A:1006150506789
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DOI: https://doi.org/10.1023/A:1006150506789