Urokinase (u-PA) and the u-PA Receptor

Modulation of in Vitro Invasiveness of Human Bladder Cancer Cell Lines
  • M’Liss A. Hudson
  • L. Michelle McReynold
Part of the Advances in Experimental Medicine and Biology book series (AEMB, volume 462)



Urokinase (u-PA) and the u-PA receptor (u-PAR) influence tumor invasion and metastasis.


The purpose of this study is to determine whether u-PAR display in 4 human bladder cancer cell lines (RT4, 253J, EJ and T24) can be modulated with substances including phorbol 12-myristate 13-acetate, interferon gamma, epidermal growth factor, and transforming growth factor beta and to correlate changes with u-PAR expression with the ability of these cells to invade artificial basement membrane (Matrigel).


u-PAR display was determined using flow cytometry and immunohistochemical staining with an anti-u-PAR monoclonal antibody (Mab3936). Matrigel invasion chamber assays were used to assess the invasive capacity of the cell lines.


The T24, EJ and 253J cells expressed the u-PAR while the RT4 cells did not. The EJ cells (expressing the highest u-PA antigen levels and the u-PAR) invaded Matrigel. The T24 cells, which expressed the u-PAR but do not produce u-PA, invaded Matrigel only when pretreated with high molecular weight urokinase (HMW u-PA). HMW u-PA pretreatment of EJ and 253J cells also enhanced their invasive potential. Blocking u-PA/u-PAR attachment with an anti-u-PAR monoclonal antibody (Mab3936) inhibited invasion. Modulation of u-PAR expression on cell lines displaying the u-PAR directly affected in vitro invasiveness of the cell lines. The RT4 cells which lack the u-PAR were not invasive under conditions tested. Thus, bladder cancer cell lines require both u-PA and the u-PAR to invade Matrigel and modulation of u-PAR display directly affects their in vitro invasive capacity.


This study shows that bladder tumor cells produce u-PA and express the u-PAR and require both for in vitro invasion to occur. When bladder cancer cells express the u-PAR, invasiveness can be enhanced by exogenous u-PA and inhibited by anti-u-PAR antibodies. Modulation of u-PAR expression on the cell surface of bladder cancer cells can also affect their ability to invade Matrigel.


Histologically similar bladder tumors show differences in their propensity to invade locally or metastasize. Intrinsic differences in the tumor cells such as the production of u-PA antigen and u-PA receptor on the cell surface and extrinsic differences in the tumor cell environment such as substances influencing u-PAR display or antibodies blocking the u-PAR may affect the biological potential of human bladder cancers and offer one explanation for the aggressive or indolent tumor behavior observed in individual patients.


Bladder Cancer Cell 253J Cell Bladder Cancer Cell Line Matrigel Invasion Assay Matrigel Invasion Chamber 
These keywords were added by machine and not by the authors. This process is experimental and the keywords may be updated as the learning algorithm improves.


Unable to display preview. Download preview PDF.

Unable to display preview. Download preview PDF.


  1. 1.
    Conese M., Blasi F.: The urokinase/urokinase-receptor system and cancer invasion. Baillieres Clin. Haematol., 8: 365, 1995.PubMedCrossRefGoogle Scholar
  2. 2.
    Cajot J.F., Kruitfof E.K., Schleuning W.D., Sordat B., Bachmann F.: Plasminogen activators, plasminogen activator inhibitors and procoagulant analyzed in twenty human tumor cell lines. Int. J. Cancer; 38: 719, 1986.PubMedCrossRefGoogle Scholar
  3. 3.
    Nykjaer A., Kjoller L., Cohen R.L., Lawrence D.A., Garni-Wagner B.A., Todd R.F. 3rd, et al.: Regions involved in binding of urokinase-type-1 inhibitor complex and pro-urokinase to endocytic alpha 2-macro-globulin receptor/low density lipoprotein receptor-related protein. Evidence that the urokinase receptor protects pro-urokinase against binding to the endocytic receptor. J. Biol. Chem., 269: 25668, 1994.;PubMedGoogle Scholar
  4. 4.
    Blasi F.: Urokinase and urokinase receptor: a paracrine/autocrine system regulating cell migration and invasiveness. Bioessays, 15: 105, 1993.PubMedCrossRefGoogle Scholar
  5. 5.
    Mignotti P., Rifkin D.B.: Biology and biochemistry of proteinase tumor invasion. Physiol. Rev. 73: 161, 1993.Google Scholar
  6. 6.
    Mizukami I.F., Garni-Wagner B.A., DeAngelo L.M., Liebert M., Flint A., Lawrence D.A., et al.: Immunologic detection of the cellular receptor for the urokinase plasminogen activator. Clin. Immunol. Immu-nopathol., 71: 96, 1994.CrossRefGoogle Scholar
  7. 7.
    Bianchi E., Cohen R.I., Thor A.T., Todd R.F. 3rd, Mizukami I.F., Lawrence D.A., et al.: The urokinase receptor is expressed in invasive breast cancer but not in normal breast tissue. Cancer Res., 54: 861, 1994.PubMedGoogle Scholar
  8. 8.
    Carnero M.V., Franco P., Del Vecchio S., Massa O., Botti G., D’Ainto G., et al.: Tissue distribution of soluble and receptor-bound urokinase in human breast cancer using a panel of monoclonal antibodies. Cancer Res., 54: 5445, 1994.Google Scholar
  9. 9.
    de Vries T.J., Quax PH., Denijn M., Verrijp K.N., Verheijen J.H., Verspaget H.W., et al.: Plasminogen activators, their inhibitors, and urokinase receptor emerge in late stages of melanocytic tumor progression. Am. J. Pathol., 144: 70, 1994.PubMedGoogle Scholar
  10. 10.
    Pyke C., Ralfkiaer E., Ronne E., Hoyer-Hansen G., Kirkeby L., Dano K.: Immunohistochemical detection of the receptor for urokinase plasminogen activator in human colon cancer. Histopathology, 24: 131, 1994.PubMedCrossRefGoogle Scholar
  11. 11.
    Stahl A., Mueller B.M.: Binding of urokinase to its receptor promotes migration and invasion of human melanoma cells in vitro.: Cancer Res., 54: 3066, 1994.PubMedGoogle Scholar
  12. 12.
    Kariko K., Kuo A., Boyd D., Okada S.S., Cines D.B., Barnathan E.S.: Overexpression of a urokinase receptor increases matrix invasion without altering cell migration in a human osteosarcoma cell line. Cancer Res., 53: 3109, 1993.PubMedGoogle Scholar
  13. 13.
    Gyetko M.R., Todd R.F. 3rd, Wilkinson C.C., Sitrin R.G.: The urokinase receptor is required for human monocyte Chemotaxis in vitro. J. Clin. Invest. 93: 1380, 1994.PubMedCrossRefGoogle Scholar
  14. 14.
    Pepper M.A., Sappino A.P., Stocklin R., Montesano R., Orci L., Vassalli, J.D.: Upregulation of urokinase receptor expression on migrating endothelial cells. J Cell. Biol., 122: 673, 1993.PubMedCrossRefGoogle Scholar
  15. 15.
    Dubeau L., Jones P.A., Rideout W.M. 3rd, Laug W.E.: Differential regulation of plasminogen activators by epidermal growth factor in normal and neoplastic human urothelium. Cancer Res., 48: 5552, 1988.PubMedGoogle Scholar
  16. 16.
    Brunner G., Pohl J., Erkell L.J., Radler-Pohl A., Schirrmacher V.: Induction of urokinase activity and malignant phenotype in bladder carcinoma cells after transfection of the activated Ha-ras oncogene. J. Cancer Res. Clin. Oncol., 115: 139, 1989.PubMedCrossRefGoogle Scholar
  17. 17.
    Hasui Y., Suzumiya J., Marutsuka K., Sumiyoshi A., Hashida S., Ishikawa E.: Comparative study of plasminogen activators in cancers and normal mucosae of human urinary bladder. Cancer Res., 49: 1067, 1989.PubMedGoogle Scholar
  18. 18.
    Hasui Y., Marutsuka K., Suzumiya J., Kitada S., Osada Y., Sumiyoshi A.: The content of urokinse-type plasminogen activator antigen as a prognostic factor in urinary bladder cancer. Int. J. Cancer, 50: 871, 1992.PubMedCrossRefGoogle Scholar
  19. 19.
    Hasui Y., Marutsuka K., Nishi S., Kitada S., Osada Y., Sumiyoshi A.: The content of urokinase-type plasminogen activator and tumor recurrence in superficial bladder cancer. J Urol., 151: 16, 1994.PubMedGoogle Scholar
  20. 20.
    Hisazumi H., Andersson L., Collins V.P.: Fibrinolytic activity of in vitro cultivated human bladder cancer cell lines. Urol. Res., 5: 133, 1977.PubMedCrossRefGoogle Scholar
  21. 21.
    Dickinson A.J., Savage P.B., Newcomb P.V., Lodge R., Sibley G.N.: The expression of urokinse, its receptor and plasminogen activator inhibitor-1 in bladder cancer [abstract]. J. Urol., 153: 406A, 1995.Google Scholar
  22. 22.
    Hudson M.A., McReynolds L.M.: Urokinase and the urokinase receptor: association with in vitro invasiveness of human bladder cancer cell lines. J. Natl. Cancer Instit., 89: 708, 1997.Google Scholar
  23. 23.
    Reuning U., Bang Nu.: Regulation of the urokinase-type plasminogen activator receptor on vascular smooth muscle cells is under the control of thrombin and other mitogens. Arterioscler. Thromb., (AZI) 12: 1161, 1992.CrossRefGoogle Scholar
  24. 24.
    Matsumoto H., Ueshima S., Fukao H., Mitsui Y., Matsuo O.: Identification of urokinase-type plasminogen activator receptor in human endothelial cells and its modulation by phorbol myristate acetate. Cell Struct. Funct., 20: 429, 1995.PubMedCrossRefGoogle Scholar
  25. 25.
    Sitrin R.G., Todd R.F. 3rd, Mizukami I.F., Gross T.J., Shollenberger S.B., Gyetko, M.R: Cytokine-specific regulation of urokinase receptor (CD87) expression by U937 mononuclear phagocytes.: Blood, 84: 1268, 1994.PubMedGoogle Scholar
  26. 26.
    Rigby C.C., Franks L.M.: A human tissue culture cell line from a transitional cell tumour of the urinary bladder: growth, chromosome pattern and ultrastructure. Br. J. Cancer, 24: 746, 1970.PubMedCrossRefGoogle Scholar
  27. 27.
    Bubenik J., Baresova M., Viklicky V., Jakoubkova J., Sainerova H., Donner J.: Established cell line of urinary bidder carcinoma (T24) containing tumor-specific antigen. Int, J. Cancer., 11: 765, 1973.CrossRefGoogle Scholar
  28. 28.
    Elliott A.Y., Cleveland P., Cervenka J., Castro A.E., Stein N., Hakala, T.R., et al.: Characterization of a cell line from human transitonal cell cancer of the urinary tract. J. Natl. Cancer Inst., 53: 1341, 1975.Google Scholar
  29. 29.
    Evans D.R., Irwin R.J., Havre P.A., Bouchard J.G., Kato T., Prout G.R., Jr.: The activity of the pyrimidine biosynthetic pathway in MGH-U1 transtional carcninoma cells grown in tissue culture. J. Urol., 127: 712, 1977.Google Scholar
  30. 30.
    Hastings R.J., Franks L.M.: Cellular heterogeneity in a tissue culture cell line derived from a human bladder carcinoma. Br. J. Cancer 47: 233, 1983.PubMedCrossRefGoogle Scholar
  31. 31.
    Chucholowski N., Schmitt M., Rettenberger.: Flow cytometric analysis of the urokinse receptor (uPAR) on tumor cells by fluorescent uPA-ligand or monoclonal antibody#3936. Fibrinolysis, 6: 95, 1992.Google Scholar
  32. 32.
    Blasi F., Stoppelli M.P., Cubellis M.V.: The receptor for urokinase-plasminogen activator. J. Cell Biochem., 32: 179, 1986.PubMedCrossRefGoogle Scholar
  33. 33.
    Hsu S.M., Raine L., Fanger H.: Use of avidin-biotin-peroxide complex (ABC) in immunoperoxidase techniques: a comparison between ABC and unlabelled antibody (PAP) procedures. J. Histochem. Cytochem., 29: 577, 1981.PubMedCrossRefGoogle Scholar
  34. 34.
    Pyke C., Kristensen P., Ralfkiaer E., Grondahl-Hansen J., Eriksen J., Blasi F., et al.: Urokinase-type plasminogen activator is expressed in stromal cells and its receptor in cancer cells at invasive foci in human colon adenocarcinomas. Am. J. Pathol., 138: 1059, 1991.PubMedGoogle Scholar
  35. 35.
    Ellis V., Pyke C., Eriksen J., Solberg H., Dano K.: The urokinase receptor: involvement in cell surface proteolysis and cancer invasion. Ann. N.Y. Acad. Sci., 667: 13, 1992.PubMedCrossRefGoogle Scholar
  36. 36.
    Dano K., Grondahl-Hansen J., Eriksen J., Nielsen B.S., Romer J., Pyke C.: The receptor for urokinase plasminogen activator:stromal cell involvement in extracellular proteolysis during cancer invasion. In: Bond JS, Barrett A J, editors. Proteolysis and protein turnover. London: Portland Press,. p239, 1993.Google Scholar
  37. 37.
    Pedersen H., Brunner N., Francis D., Osterlind K., Ronne E., Hansen H.H., et al.: Prognostic impact of urokinase, urokinase receptor, and type 1 plasminogen activator inhibitor in squamous and large cell lung cancer tissue. Cancer Res., 54: 4671, 1994.PubMedGoogle Scholar
  38. 38.
    Reiter L.S., Kruithof E.K., Cajot J., Sordat B.: The role of the urokinase receptor in extracellular matrix degradation by HT29 human colon carinoma cells. Int J. Cancer, 53: 444, 1993.PubMedCrossRefGoogle Scholar
  39. 39.
    Redwood S.M., Liu B.C., Weiss R.E., Hodge D.E., Droller M.J.: Abrogation of the invasion of human bladder tumor cells by using protease inhibitor(s). Cancer, 69: 1212, 1992.PubMedCrossRefGoogle Scholar
  40. 40.
    Ossowski L., Russo-Payne H., Wilson E.L.: Inhibition of urokinase-type plasminogen activator by antibodies: the effect on dissemination of a human tumor in the nude mouse. Cancer Res., 51: 274, 1991.PubMedGoogle Scholar
  41. 41.
    Ossowski L.: Invasion of connective tissue by human carcinoma cell lines: requirment for urokinase, urokinse receptor, and interstitial collagenase. Cancer Res., 52: 6754, 1992.PubMedGoogle Scholar
  42. 42.
    Hearing V.J., Law L.W., Conti A., Apella E., Blasi F.: Modulation of metastatic potential by cell surface urokinase of murine cells. Cancer Res., 48: 1270, 1988.PubMedGoogle Scholar
  43. 43.
    Crowley C.W., Cohen R.L., Lucas B.K., Liu G., Shuman M.A., Levinson A.D.: Prevention of metastasis by inhibition of the urokinase receptor. Proc. Natl. Acad. Sci. U.S.A., 90: 5021, 1993.PubMedCrossRefGoogle Scholar
  44. 44.
    Schuster W.A., Medcalf R.L., Kruithof E.K.: Retinoic acid potentiates phorbol ester-mediated induction of urokinase and plasminogen activator inhibitor type 2 in human myeloid leukemic cell lines. Endocrinology, 133: 1724, 1993.PubMedCrossRefGoogle Scholar
  45. 45.
    Hansen S.H., Behrendt N., Dano K., Kristensen P.: Localization of urokinase-type plasminogen activator receptor on U937 cells: phorbol ester PMA induces heterogeneity. Exp. Cell. Res., 187: 255, 1990.PubMedCrossRefGoogle Scholar

Copyright information

© Springer Science+Business Media New York 1999

Authors and Affiliations

  • M’Liss A. Hudson
    • 1
  • L. Michelle McReynold
    • 1
  1. 1.Division of Urologic SurgeryWashington University School of MedicineSt. LouisUSA

Personalised recommendations