Vitronectin binding to urokinase receptor in human breast cancer

Maria V. Carriero, Silvana Del Vecchio, Paola Franco, Maria I. Potena, Ferdinando Chiaradonna, Gerardo Botti, M. Patrizia Stoppelli, Marco Salvatore

Research output: Contribution to journalArticlepeer-review

Abstract

Functional assembly of the plasminogen-dependent proteolytic system on the cell surface requires multiple interactions involving urokinase (uPA), urokinase receptor (uPAR), plasminogen activator inhibitors, and other molecules that mediate cell migration and adhesion. We analyzed the in vitro interaction of uPAR-containing particulate cell fractions with the amino- terminal fragment (ATF) of human urokinase and the matrix-like form of vitronectin. Binding and cross-linking of 125I-labeled ATF to crude membrane extracts from LB6-19 mouse cells overexpressing human uPARs in the presence of 25 nM urea-denatured vitronectin led to the formation of M(r) 137,000, 92,000, and 82,000 covalent complexes. Immunoprecipitation of the preformed cross-linked 125I-labeled complexes with anti-vitronectin, anti- uPA, or anti-uPAR antibodies revealed that the M(r) 82,000 and 92,000 species do contain ATF and vitronectin and identified the M(r) 137,000 species as a ternary complex formed by ATF, uPAR, and vitronectin. A similar electrophoretic pattern was displayed by acid-pretreated membranes extracted from MCF-7 breast carcinoma or HT1080 fibrosarcoma cell lines, as well as a ductal breast carcinoma specimen; the latter exhibited complex formation at concentrations of vitronectin lower than 10 nM. Finally uPAR-vitronectin interaction was further documented by the decreased reactivity of an anti- uPAR polyclonal antibody to acid-pretreated sections of 10 breast carcinomas that had been preincubated with vitronectin. Our findings highlight the ability of uPAR to interact simultaneously with vitronectin and uPA in breast cancer, supporting a dynamic coupling of the molecular mechanisms underlying plasminogen-dependent matrix degradation and cell adhesion.

Original languageEnglish
Pages (from-to)1299-1308
Number of pages10
JournalClinical Cancer Research
Volume3
Issue number8
Publication statusPublished - Aug 1997

ASJC Scopus subject areas

  • Cancer Research
  • Oncology

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