Quantitative assessment of interaction of urokinase-type plasminogen activator and its receptor (CD87) by use of a solid-phase uPA-ligand binding assay

Goretzki, Lothar; Bognacki, John; Koppitz, Marcus; Rettenberger, P.; Magdolen, Viktor; Creutzburg, Sabine; Hammelburger, J W; Weidle, Ulrich H.; Wilhelm, Olaf G.; Kessler, Horst; Graeff, H.

doi:10.1016/s0268-9499(97)80004-5

Cited by 9 publications

(4 citation statements)

References 26 publications

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“…Applying a solid-phase uPA ligand-binding assay (Goretzki et al 1997), we proved that MDA-MB-suPAR cells produced and secreted suPAR as an active, uPA-binding receptor. In contrast to the findings with stably transfected ovarian cancer cells, high-level synthesis of suPAR did not alter the cell doubling rates compared to the parental cell line.…”

Section: In Vitro and In Vivo Effects Of High-level Synthesis Of Recomentioning

confidence: 92%

Inhibition of the Tumor-Associated Urokinase-Type Plasminogen Activation System: Effects of High-Level Synthesis of Soluble Urokinase Receptor in Ovarian and Breast Cancer Cells in Vitro and in Vivo

Magdolen

Krüger²,

Sato³

et al. 2003

Molecular Staging of Cancer

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Tumor cell invasion and metastasis depend on the coordinated and temporal expression of proteolytic enzymes to degrade the surrounding extracellular matrix and of adhesion molecules to remodel cell-cell and/or cell-matrix attachments. The tumor cell-associated urokinase-type plasminogen activator system, consisting of the serine protease uPA, its substrate plasminogen, its membrane-bound receptor uPAR, as well as its inhibitors PAI-l and PAI-2, plays an important role in these pericellular processes. Especially, association of the proteolytic activity of uPA with the cell surface via interaction with uPAR significantly increases the invasive capacity of tumor cells. Consequently, various approaches have been pursued to interfere with the expression or activity of uPA and/or uPAR, including antisense strategies and the development of active-site inhibitors of uPA or inhibitors of uPA/uPAR interaction. In this review, we focus on the results obtained in vitro and in vivo with tumor cells producing high levels of a recombinant soluble form of uPAR, which efficiently inhibits uPA binding to cell surface-associated uPAR and, by this, acts as a scavenger for uPA.

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Section: In Vitro and In Vivo Effects Of High-level Synthesis Of Recomentioning

confidence: 92%

Inhibition of the Tumor-Associated Urokinase-Type Plasminogen Activation System: Effects of High-Level Synthesis of Soluble Urokinase Receptor in Ovarian and Breast Cancer Cells in Vitro and in Vivo

Magdolen

Krüger²,

Sato³

et al. 2003

Molecular Staging of Cancer

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“…uPA, PAI‐1, uPAR, and D‐dimer concentrations were determined by ELISA kits Imubind uPA # 894, Imubind PAI‐1 # 821, Imubind uPAR # 893, and D‐dimer ELISA, respectively (American Diagnostica). Solid phase uPA ligand binding assays were performed as published [16]. Recombinant soluble uPAR (uPAR 1–277 ) expressed in Chinese hamster ovary (CHO) cells was purified from cell culture supernatants by uPA ligand affinity chromatography and its uPA binding activity assessed by flow cytofluorometry [17].…”

Section: Methodsmentioning

confidence: 99%

Urokinase induces proliferation of human ovarian cancer cells: characterization of structural elements required for growth factor function

et al. 1998

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Ovarian cancer metastasis is associated with an increase in the urokinase-type plasminogen activator (uPA) and its receptor uPAR. We present evidence that binding of uPA to uPAR provokes a mitogenic response in the human ovarian cancer cell line OV-MZ-6 in which endogenous uPA production had been significantly reduced by stable uPA`antisense' transfection. High molecular weight (HMW) uPA, independent of its enzymatic activity, produced an up to 95% increase in cell number concomitant with 2-fold elevated [Q H]thymidine incorporation as did the catalytically inactive but uPAR binding amino-terminal fragment of uPA, ATF. uPA-induced cell proliferation was significantly decreased by blocking uPA/uPAR interaction by the monoclonal antibody IIIF10 and by soluble uPAR. The efficiency of the uPAR binding synthetic peptide cyclo IWYQI uPA IWÀQI to enhance OV-MZ-6 cell growth proved this molecular domain to be the minimal structural determinant for uPA mitogenic activity. Dependence of uPA-provoked cell proliferation on uPAR was further demonstrated in Raji cells which do not express uPAR and were thus not induced by uPA. However, upon transfection with full-length uPAR, Raji cells acquired a significant growth response to HMW uPA and ATF.z 1998 Federation of European Biochemical Societies.

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“…The solid-phase uPA ligand-binding assay was performed in a similar manner as described previously [21]. Briefly, wells of microtiter plates (Maxisorb; Nunc) were coated overnight at 4 o C with 100 µl of rec-uPAR (1 µg/ml).…”

Section: Solid-phase Upa Binding Assaymentioning

confidence: 99%

Overexpression of the urokinase receptor mRNA splice variant uPAR-del4/5 affects tumor-associated processes of breast cancer cells in vitro and in vivo

Sato

Kopitz²,

Grismayer

et al. 2010

Breast Cancer Res Treat

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Quantitative assessment of interaction of urokinase-type plasminogen activator and its receptor (CD87) by use of a solid-phase uPA-ligand binding assay

Cited by 9 publications

References 26 publications

Inhibition of the Tumor-Associated Urokinase-Type Plasminogen Activation System: Effects of High-Level Synthesis of Soluble Urokinase Receptor in Ovarian and Breast Cancer Cells in Vitro and in Vivo

Inhibition of the Tumor-Associated Urokinase-Type Plasminogen Activation System: Effects of High-Level Synthesis of Soluble Urokinase Receptor in Ovarian and Breast Cancer Cells in Vitro and in Vivo

Urokinase induces proliferation of human ovarian cancer cells: characterization of structural elements required for growth factor function

Overexpression of the urokinase receptor mRNA splice variant uPAR-del4/5 affects tumor-associated processes of breast cancer cells in vitro and in vivo

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