Abstract
A recombinant soluble human urokinase receptor comprising amino acids 1-277 was cloned and transfected into CHO cells. The mutant protein (rec-uPAR277), purified from the CHO cell supernatant by affinity chromatography on immobilized urokinase (uPA), in a four-fold excess, completely abolished the binding of FITC-labeled pro-uPA to the human ovarian cancer cell line, OV-MZ-6. This invasive and tumorigenic cancer cell line expresses uPA, its inhibitor PAI-1, and the high-affinity receptor for uPA, uPAR. Rec-uPAR277 significantly reduced the proliferation of OV-MZ-6 cells in a concentration-dependent manner without altering the viability of the cells. Invasion of OV-MZ-6 cells tested in an in vitro Matrigel invasion assay was inhibited by rec-uPAR277 up to 75%. In conclusion, these results demonstrate that rec-uPAR277 can function as a scavenger for uPA in vitro by inhibiting proliferation and invasion of human cancer cells.
Publication types
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Research Support, Non-U.S. Gov't
MeSH terms
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Animals
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CHO Cells
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Cell Division
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Cell Line
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Cell Survival
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Chromatography, Affinity
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Cloning, Molecular
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Cricetinae
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Cystadenocarcinoma / metabolism
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Cystadenocarcinoma / pathology
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Female
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Humans
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Mutagenesis, Site-Directed
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Neoplasm Invasiveness
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Ovarian Neoplasms / metabolism
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Ovarian Neoplasms / pathology
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Plasminogen Activator Inhibitor 1 / biosynthesis
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Receptors, Cell Surface / biosynthesis
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Receptors, Cell Surface / isolation & purification
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Receptors, Cell Surface / metabolism*
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Receptors, Urokinase Plasminogen Activator
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Recombinant Proteins / biosynthesis
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Recombinant Proteins / isolation & purification
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Recombinant Proteins / metabolism
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Transfection
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Tumor Cells, Cultured
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Urokinase-Type Plasminogen Activator / metabolism*
Substances
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PLAUR protein, human
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Plasminogen Activator Inhibitor 1
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Receptors, Cell Surface
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Receptors, Urokinase Plasminogen Activator
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Recombinant Proteins
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Urokinase-Type Plasminogen Activator