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[Cancer Research 61, 278-284, January 1, 2001]
© 2001 American Association for Cancer Research

Immunology

Mutant E-cadherin Breast Cancer Cells Do Not Display Constitutive Wnt Signaling1

Marc van de Wetering, Nick Barker, I. Clara Harkes, Marcel van der Heyden, Nicolette J. Dijk, Antoinette Hollestelle, Jan G. M. Klijn, Hans Clevers and Mieke Schutte2

Department of Immunology [M. v. d. W., N. B., H. C.] and Center for Biomedical Genetics [H. C.], University Medical Center Utrecht, 3508 GA Utrecht, the Netherlands; Department of Medical Oncology, Josephine Nefkens Institute, Erasmus Medical Center Rotterdam, 3000 DR Rotterdam, the Netherlands [I. C. H., N. J. D., A. H., J. G. M. K., M. S.]; and Hubrecht Laboratory, Netherlands Institute for Developmental Biology, 3584 GT Utrecht, the Netherlands [M. v. d. H.]

Participation of E-cadherin in the Wnt signaling pathway was suggested because of the dual role of ß-catenin in cell adhesion and the Wnt signaling cascade. Whereas ß-catenin interacts at the cell membrane with the cell adhesion protein E-cadherin, in the nucleus it activates Wnt target genes through formation of transcriptionally active complexes with members of the Tcf/Lef family of transcription factors. Here, we analyzed by PCR and direct cycle sequencing 26 human breast cancer cell lines for alterations in the E-cadherin gene. Genetic alterations were identified in eight cell lines. Five cell lines had truncating mutations, whereas three cell lines had in-frame deletions in the gene transcript and expressed mutant E-cadherin proteins at the cell membrane. Involvement of E-cadherin in the Wnt pathway was evaluated through determination of the activity of a Tcf reporter gene, which had been transiently transfected into 15 breast cancer cell lines. None of six E-cadherin mutant cell lines and four cell lines that exhibit transcriptional silencing of the E-cadherin gene showed Tcf-mediated transcriptional activation. E-cadherin wild-type cell line DU4475 exhibited constitutive Tcf-ß-catenin signaling activity and was found to express truncated APC proteins. These results indicate that if cellular transformation occurred through mutation of E-cadherin, it is not mediated via constitutive activation of the Wnt signaling pathway.




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Copyright © 2001 by the American Association for Cancer Research.