Urokinase induces receptor mediated brain tumor cell migration and invasion

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.

[1]  B. M. Mueller,et al.  Binding of urokinase to its receptor promotes migration and invasion of human melanoma cells in vitro. , 1994, Cancer research.

[2]  D. Waltz,et al.  Reversible cellular adhesion to vitronectin linked to urokinase receptor occupancy. , 1994, The Journal of biological chemistry.

[3]  P. Quax,et al.  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 , 1991, The Journal of cell biology.

[4]  M. Ploug,et al.  Protein Structure and Membrane Anchorage of the Cellular Receptor for Urokinase-Type Plasminogen Activator , 1991, Seminars in thrombosis and hemostasis.

[5]  P. Jones,et al.  Destruction of extracellular matrices containing glycoproteins, elastin, and collagen by metastatic human tumor cells. , 1980, Cancer research.

[6]  D. Belin,et al.  A cellular binding site for the Mr 55,000 form of the human plasminogen activator, urokinase , 1985, The Journal of cell biology.

[7]  G. Nenci,et al.  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 , 1996, Cancer.

[8]  Z. Gokaslan,et al.  In vitro inhibition of human glioblastoma cell line invasiveness by antisense uPA receptor , 1997, Oncogene.

[9]  Michael V. Doyle,et al.  Regulation of Integrin Function by the Urokinase Receptor , 1996, Science.

[10]  P. Verde,et al.  Urokinase-dependent cell surface proteolysis and cancer. , 1990, Seminars in cancer biology.

[11]  D. Rifkin,et al.  Tumor invasion through the human amniotic membrane: Requirement for a proteinase cascade , 1986, Cell.

[12]  A. Levinson,et al.  Prevention of metastasis by inhibition of the urokinase receptor. , 1993, Proceedings of the National Academy of Sciences of the United States of America.

[13]  O. Majdic,et al.  Urokinase plasminogen activator receptor, beta 2-integrins, and Src- kinases within a single receptor complex of human monocytes , 1995, The Journal of experimental medicine.

[14]  D. Cines,et al.  Overexpression of urokinase receptor increases matrix invasion without altering cell migration in a human osteosarcoma cell line. , 1993, Cancer research.

[15]  W. Paulus,et al.  Basement membrane invasion of glioma cells mediated by integrin receptors. , 1994, Journal of neurosurgery.

[16]  Y. Kook,et al.  The effect of antisense inhibition of urokinase receptor in human squamous cell carcinoma on malignancy. , 1994, The EMBO journal.

[17]  L. Zanetta,et al.  Control of type IV collagenase activity by components of the urokinase–plasmin system: a regulatory mechanism with cell‐bound reactants , 1997, The EMBO journal.

[18]  P. Phillips,et al.  Malignant gliomas in children. , 1996, Cancer investigation.

[19]  M. Berens,et al.  Determinants of human astrocytoma migration. , 1994, Cancer research.

[20]  K. Danø,et al.  Plasminogen activators, tissue degradation, and cancer. , 1985, Advances in cancer research.

[21]  I. Pollack Brain tumors in children. , 1994, The New England journal of medicine.

[22]  S. Waxman,et al.  Induction of cell migration by pro-urokinase binding to its receptor: possible mechanism for signal transduction in human epithelial cells , 1994, The Journal of cell biology.

[23]  J. Furuyama,et al.  Matrix metalloproteinases and tissue inhibitors of metalloproteinases in human gliomas. , 1995, Journal of neurosurgery.

[24]  S. Kohga,et al.  Localization of plasminogen activators in human colon cancer by immunoperoxidase staining. , 1985, Cancer research.

[25]  P. Gudewicz,et al.  Human urokinase-type plasminogen activator stimulates chemotaxis of human neutrophils. , 1987, Biochemical and biophysical research communications.

[26]  Andreas S. Beutler,et al.  Diffuse brain invasion of glioma cells requires β1 integrins , 1996 .

[27]  A. Vaheri,et al.  Directed plasminogen activation at the surface of normal and malignant cells. , 1991, Advances in cancer research.

[28]  D. Rifkin,et al.  Urokinase‐type plasminogen activator mediates basic fibroblast growth factor‐induced bovine endothelial cell migration independent of its proteolytic activity , 1992, Journal of cellular physiology.

[29]  N. Brünner,et al.  Prognostic impact of urokinase, urokinase receptor, and type 1 plasminogen activator inhibitor in squamous and large cell lung cancer tissue. , 1994, Cancer research.

[30]  S. Rosenberg,et al.  Identification of the urokinase receptor as an adhesion receptor for vitronectin. , 1994, The Journal of biological chemistry.

[31]  H. Yoshioka,et al.  Extracellular matrix production and degradation by adenoid cystic carcinoma cells: participation of plasminogen activator and its inhibitor in matrix degradation. , 1993, Cancer research.

[32]  Repesh La A new in vitro assay for quantitating tumor cell invasion. , 1989 .

[33]  L. Magnelli,et al.  Role of specific membrane receptors in urokinase-dependent migration of human keratinocytes. , 1990, The Journal of investigative dermatology.

[34]  W. Laug,et al.  Clonal variation of expression of the genes coding for plasminogen activators, their inhibitors and the urokinase receptor in HT1080 sarcoma cells , 1992, International journal of cancer.

[35]  J. Bruner,et al.  Expression and localization of urokinase-type plasminogen activator in human astrocytomas in vivo. , 1994, Cancer research.

[36]  W. Laug,et al.  Endothelial Cells Degrade Extracellular Matrix Proteins Produced In Vitro , 1985, Thrombosis and Haemostasis.

[37]  M. Olman,et al.  Up-regulation of urokinase and urokinase receptor genes in malignant astrocytoma. , 1995, The American journal of pathology.

[38]  E. Appella,et al.  The receptor-binding sequence of urokinase. A biological function for the growth-factor module of proteases. , 1987, The Journal of biological chemistry.