NF-κB is essential for epithelial-mesenchymal transition and metastasis in a model of breast cancer progression

Margit Huber

Ninel Azoitei

Bernd Baumann

Stefan Grünert

^{Department of Physiological Chemistry, Ulm University, Ulm, Germany.}^{Institute of Molecular Pathology, Vienna, Austria.}^{Department of Dermatology, Vienna Medical University, Vienna, Austria.}^{Department of New Chemical Entity Lead Discovery, Boehringer Ingelheim Austria GmbH, Vienna, Austria.}

Address correspondence to: Thomas Wirth, Department of Physiological Chemistry, Ulm University, Albert-Einstein-Allee 11, D-89081 Ulm, Germany. Phone: 49-731-500-23270; Fax: 49-731-500-22892; E-mail: ed.mlu-inu.nizidem@htriw.samoht.

Received 2004 Feb 17; Accepted 2004 Jun 22.

Abstract

The transcription factor NF-κB is activated in a range of human cancers and is thought to promote tumorigenesis, mainly due to its ability to protect transformed cells from apoptosis. To investigate the role of NF-κB in epithelial plasticity and metastasis, we utilized a well-characterized in vitro/in vivo model of mammary carcinogenesis that depends on the collaboration of the Ha-Ras oncoprotein and TGF-β. We show here that the IKK-2/IκBα/NF-κB pathway is required for the induction and maintenance of epithelial-mesenchymal transition (EMT). Inhibition of NF-κB signaling prevented EMT in Ras-transformed epithelial cells, while activation of this pathway promoted the transition to a mesenchymal phenotype even in the absence of TGF-β. Furthermore, inhibition of NF-κB activity in mesenchymal cells caused a reversal of EMT, suggesting that NF-κB is essential for both the induction and maintenance of EMT. In line with the importance of EMT for invasion, blocking of NF-κB activity abrogated the metastatic potential of mammary epithelial cells in a mouse model system. Collectively, these data provide evidence of an essential role for NF-κB during distinct steps of breast cancer progression and suggest that the cooperation of Ras- and TGF-β–dependent signaling pathways in late-stage tumorigenesis depends critically on NF-κB activity.

Abstract

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Acknowledgments

We are grateful to M. Jechlinger for providing EpRas and EpRasXT cells; B. Anic and G. Litos for excellent technical assistance; K. Stangl and P. Garin-Chesa for help with histology; the Institute of Molecular Pathology BioOptics Department for support with immunofluorescence microscopy; and K. Scharffetter-Kochanek, P. Petzlbauer, C. Hoeller, S. Maschler, and M. Herlyn for scientific discussions and useful comments. This work was in part funded by the Genome Research in Austria (GEN-AU) program (to M.A. Huber) and by the German Science Foundation (DFG SFB497/B1 to B. Baumann; SFB451/A9 to T. Wirth).

Acknowledgments

Footnotes

Nonstandard abbreviations used: benzyloxycarbonyl-Val-Ala-Asp-fluoromethyl-ketone (Z-VAD-FMK); constitutively active (CA); electrophoretic mobility-shift assay (EMSA); epithelial-mesenchymal transition (EMT); IκB kinase (IKK); inhibitor κB (IκB); NF-κB essential modulator (NEMO); TGF-β1–activated kinase 1 (TAK1); trans-dominant (TD).

Conflict of interest: The authors have declared that no conflict of interest exists.

Footnotes

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