Determination of the Levels of Urokinase and Its Receptor in Human Colon Carcinoma Cell Lines

Douglas Boyd, Germaine Florent, Paul Kim, Michael Brattain

Research output: Contribution to journalArticlepeer-review

110 Scopus citations

Abstract

At present, there is a lack of availability of differentiation markers for colon carcinoma. This may, in part, be a consequence of the diversified function of the normal human colon. This study addresses the possibility that the expression of urokinase and its receptor is inversely related to differentiation in colon carcinoma. Six colon carcinoma cell lines including three well-differentiated (CBS, GEO, FET) and three poorly differentiated ones (HCT116, HCT116b, RKO) were screened for urokinase receptor display and secretion of the plasminogen activator. A radioreceptor assay was used to determine receptor levels. Binding of radioactive urokinase to colon cells was saturable, specific, and time dependent. Cell-bound 125I-labeled protease was unaffected by the presence of epidermal growth factor, low-molecular-weight urokinase plasminogen, or transferrin. Time course studies revealed that maximum amounts of radioactive tracer were bound in a 30-min period with no change occurring over the course of a 90-min incubation. Scatchard analysis of ligand binding indicated that the well-and poorly differentiated cells could be separated on the basis of receptor display; the aggressive RKO, HCT116, and HCT116b expressed in excess of 105 sites per cell, while the more indolent CBS, GEO, and FET possessed less than 1.5 x 104 receptors per cell. The colon carcinoma cells were also analyzed for urokinase in the conditioned medium. Low levels of the plasminogen activator (0.8 to 1.3 ng/ml/10` cells/72 h) were associated with the more “mature” cells. This was in contrast to the elevated levels of the protease (3.9 to 11.4 ng/ml/ 106 cells/72 h) present in the medium derived from the more aggressive cells (HCT 116, HCT116b, RKO). Thus, secreted urokinase and/or the expression of cellular receptor for the plasminogen activator may provide useful measurements of the degree of undifferentiation of in vitro colon carcinoma.

Original languageEnglish (US)
Pages (from-to)3112-3116
Number of pages5
JournalCancer Research
Volume48
Issue number11
StatePublished - 1988

ASJC Scopus subject areas

  • Oncology
  • Cancer Research

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