Regular Article

British Journal of Cancer (1999) 80, 1884–1891. doi:10.1038/sj.bjc.6690616 www.bjcancer.com
Published online 30 July 1999

Targeted disruption of the K-Ras oncogene in an invasive colon cancer cell line down-regulates urokinase receptor expression and plasminogen-dependent proteolysis

H Allgayer1,3, H Wang1, S Shirasawa2, T Sasazuki2 and D Boyd1

  1. 1Departments of Cancer Biology, PO Box 108, The University of Texas MD Anderson Cancer Center, 1515 Holcombe Blvd, Houston, TX 770 30, USA
  2. 2Department of Genetics, Kyushu University, Fukuoka, Japan
  3. 3Present address: Department of Surgery, Klinikum Grosshadern, Ludwig-Maximillians University, Munich, Germany

Received 17 November 1998; Revised 4 March 1999; Accepted 9 March 1999.

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Abstract

The urokinase receptor, overexpressed in invasive colon cancer, promotes tumour cell invasion. Since K-Ras is activated in many colon cancers, we determined if urokinase receptor overexpression is a consequence of this activated oncogene. Accordingly, urokinase receptor expression was compared in HCT 116 colon cancer cells containing either a mutation-activated K-Ras or disrupted for this oncogene (by homologous recombination). HCT 116 cells containing the disrupted K-Ras oncogene expressed between 50 and 85% less urokinase receptor protein compared with the parental HCT 116 cells. Reduced urokinase receptor expression in cells containing the disrupted mutated K-Ras was not due to a physical impairment of the urokinase receptor gene since phorbol ester treatment was inductive for its expression. Constitutive urokinase receptor expression in HCT 116 cells required an intact AP-1 motif in the promoter (at –184) and electrophoretic mobility shifting assays indicated less c-Jun, JunD, c-Fos and Fra-1 bound to this motif in the K-Ras-disrupted cells. Since the urokinase receptor accelerates proteolysis, laminin degradation was compared in cells containing the mutation-activated and disrupted K-Ras oncogene. The latter cells displaying fewer urokinase receptors, degraded 80% less laminin. This is the first study to demonstrate a role for K-Ras as a regulator of the constitutive expression of the urokinase receptor.

Keywords:

u-PAR, K-Ras, colon cancer, knockout, proteolysis

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