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Molecular and Cellular Biology, August 1999, p. 5696-5706, Vol. 19, No. 8
0270-7306/99/$04.00+0
Copyright © 1999, American Society for Microbiology. All rights reserved.

Neoplastic Transformation of RK3E by Mutant beta -Catenin Requires Deregulation of Tcf/Lef Transcription but Not Activation of c-myc Expression

Frank T. Kolligs,1 Gang Hu,1 Chi V. Dang,2 and Eric R. Fearon1,3,4,*

Division of Molecular Medicine & Genetics and the Cancer Center, Departments of Internal Medicine,1 Human Genetics,3 and Pathology,4 University of Michigan School of Medicine, Ann Arbor, Michigan 48109, and Department of Medicine and the Oncology Center, Johns Hopkins University School of Medicine, Baltimore, Maryland 212052

Received 8 March 1999/Returned for modification 19 April 1999/Accepted 12 May 1999

Current models predict that beta -catenin (beta -cat) functions in Wnt signaling via activation of Tcf/Lef target genes and that its abundance is regulated by the adenomatous polyposis coli (APC) and glycogen synthase kinase 3beta (GSK3beta ) proteins. In colon and other cancers, mutations in APC or presumptive GSK3beta phosphorylation sites of beta -cat are associated with constitutive activation of Tcf/Lef transcription. In spite of assumptions about its oncogenic potential, prior efforts to demonstrate that mutated beta -cat will induce neoplastic transformation have yielded equivocal results. We report here that mutated, but not wild-type, beta -cat proteins induced neoplastic transformation of RK3E, an adenovirus E1A-immortalized epithelial cell line. Analysis of the properties of mutant beta -cat proteins and studies with a dominant negative Tcf-4 mutant indicated that the ability of beta -cat to bind and activate Tcf/Lef factors is crucial for transformation. c-myc has recently been implicated as a critical Tcf-regulated target gene. However, c-myc was not consistently activated in beta -cat-transformed RK3E cells, and a dominant negative c-Myc mutant protein failed to inhibit beta -cat transformation. Our findings underscore the role of beta -cat mutations and Tcf/Lef activation in cancer and illustrate a useful system for defining critical factors in beta -cat transformation.

* Corresponding author. Mailing address: Division of Molecular Medicine & Genetics, University of Michigan Medical Center, 4301 MSRB 3, 1150 W. Medical Center Dr., Ann Arbor, MI 48109-0638. Phone: (734) 764-1549. Fax: (734) 647-7979. E-mail: fearon{at}umich.edu.

Molecular and Cellular Biology, August 1999, p. 5696-5706, Vol. 19, No. 8
0270-7306/99/$04.00+0
Copyright © 1999, American Society for Microbiology. All rights reserved.


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