Abstract
Several cell biological studies have shown that the invasiveness of a variety of tumors depend on the regulated expression of proteolytic enzymes that degrade the surrounding extracellular matrix and dissociate cell-cell and/or cell-matrix attachments. One such enzyme, the serine protease urokinase-type plasminogen activator (uPA), converts enzymatically inactive plasminogen into the widely acting protease plasmin, which degrades several extracellular matrix components and also activates proenzyme forms of matrix metalloproteases. Thus, uPA is a central molecule in pericellular proteolysis (1-1). uPA (as well as other factors of the plasminogen activator system, the cell surface-associated uPA receptor [uPAR], and the plasminogen activator inhibitor type-1 [PAI-1]) is an important prognostic factor predicting relapse-free and/or survival in patients with a variety of solid malignant tumors including ovarian cancer; in all cases, high levels of uPA are associated with a poor prognosis (4-6).
This is a preview of subscription content, log in via an institution to check access.
Access this chapter
Tax calculation will be finalised at checkout
Purchases are for personal use only
References
Dano, K., Andreasen, P. A., Grondahl-Hansen, J., Kristensen, P., Nielsen, L. S., and Skriver, L. (1985) Plasminogen activators, tissue degradation, and cancer. Adv. Cancer Res. 44, 139–266.
Schmitt, M., Wilhelm, O., Jänicke, F., Magdolen, V., Reuning, U., Ohi, H., et al. (1995) Urokinase-type plasminogen activator (uPA) and its receptor (CD87): a new target in tumor invasion and metastasis. J. Obstet. Gynaecol. 21, 151–165.
Blasi, F. (1997) uPA, uPAR, PAI-1: key intersection of proteolytic, adhesive and chemotactic highways? Immunol. Today 18, 415–417.
Schmitt, M., Harbeck, N., Thomssen, C., Wilhelm, O., Magdolen, V., Reuning, U., et al. (1997) Clinical impact of the plasminogen activation system in tumor invasion and metastasis: prognostic relevance and target for therapy. Thromb. Haemost. 78, 285-296.
Kuhn, W., Pache, L., Schmalfeldt, B., Dettmar, P., Schmitt, M., Jänicke, F., and Graeff, H. (1994) Urokinase (uPA) and PAI-1 predict survival in advanced ovarian cancer patients (FIGO III) after radical surgery and platinum-based chemotherapy. Gynecol. Oncol. 55, 401–409.
Schmalfeldt, B., Kuhn, W., Reuning, U., Pache, L., Dettmar, P., Schmitt, M., et al. (1995) Primary tumor and metastasis in ovarian cancer differ in their content of urokinase-type plasminogen activator, its receptor, and inhibitors types 1 and 2. Cancer Res. 55, 3958-3963.
Türkmen, B., Magdolen, V., Trommler, P., Creutzburg, S., Graeff, H., and Schmitt, M. (1997) Sequence analysis of cDNAs encoding P AI-1, uP A and uPAR from ovarian cancer cell lines, in Adv. Exp. Biol. 425, 266–267.
Türkmen, B., Schmitt, M., Schmalfeldt, B., Trommler, P., Hell, W., Creutzburg, S., et al. (1997) Mutational analysis of the genes encoding urokinase-type plasminogen activator (uPA) and its inhibitor (PAI-1) in advanced ovarian cancer Electrophoresis 18, 686–689.
Chomczynski, P. and Sachi, N. (1987) Single step method of RNA isolation by acid guanidinium thiocyanate-phenol-chloroform extraction. Anal. Biochem. 162, 156.
Chomczynski, P. (1995) A reagent for the single-step simultaneous isolation of RNA, DNA and proteins from cell and tissue samples. BioTechniques 15, 532–534.
Sambrook, J., Fritsch, E. F., and Maniatis, T. (1989) Molecular Cloning. A Laboratory Manual. Cold Spring Harbor Laboratory Press, Cold Spring Harbor, New York.
Rolfs, A., Schuller, I., Finckh, U., and Weber-Rolfs, I. (1992) PCR: Clinical Diagnostics and Research. Springer Laboratory, Springer-Verlag, Berlin, Germany.
Tautz, D. and Renz, M. (1993) An optimized freeze-squeeze method for the recovery of DNA fragments from agarose gels. Anal. Biochem. 132, 14–19.
Wittwer, C., Ririe, K., Andrew, R., David, D., Gundry, R., and Balis, U. (1997) The LightCycler: a micro-volume multisample fluorimeter with rapid temperature control. BioTechniques 22, 176–181.
Author information
Authors and Affiliations
Editor information
Editors and Affiliations
Rights and permissions
Copyright information
© 2000 Humana Press Inc.
About this protocol
Cite this protocol
Muehlenweg, B. et al. (2000). RFLP Molecular Analysis of the Urokinase-Type Plasminogen Activator Gene. In: Bartlett, J.M.S. (eds) Ovarian Cancer. Methods in Molecular Medicine™, vol 39. Humana Press. https://doi.org/10.1385/1-59259-071-3:299
Download citation
DOI: https://doi.org/10.1385/1-59259-071-3:299
Publisher Name: Humana Press
Print ISBN: 978-0-89603-583-6
Online ISBN: 978-1-59259-071-1
eBook Packages: Springer Protocols