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Specific interactions between Smad proteins and AP-1 components determine TGFβ-induced breast cancer cell invasion

Oncogene volume 32, pages 3606–3615 (2013)Cite this article

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Abstract

Deregulation of the transforming growth factor β (TGFβ) signal transduction cascade is functionally linked to cancer. In early phases, TGFβ acts as a tumor suppressor by inhibiting tumor cell proliferation, whereas in late phases, it can act as a tumor promoter by stimulating tumor cell invasion and metastasis. Smad transcriptional effectors mediate TGFβ responses, but relatively little is known about the Smad-containing complexes that are important for epithelial–mesenchymal transition and invasion. In this study, we have tested the hypothesis that specific members of the AP-1 transcription factor family determine TGFβ signaling specificity in breast cancer cell invasion. Using a 3D model of collagen-embedded spheroids of MCF10A-MII premalignant human breast cancer cells, we identified the AP-1 transcription factor components c-Jun, JunB, c-Fos and Fra1 as essential factors for TGFβ-induced invasion and found that various mesenchymal and invasion-associated TGFβ-induced genes are co-regulated by these proteins. In situ proximity ligation assays showed that TGFβ signaling not only induces complexes between Smad3 and Smad4 in the nucleus but also complexes between Smad2/3 and Fra1, whereas complexes between Smad3, c-Jun and JunB could already be detected before TGFβ stimulation. Finally, chromatin immunoprecipitations showed that c-Jun, JunB and Fra1, but not c-Fos, are required for TGFβ-induced binding of Smad2/3 to the mmp-10 and pai-1 promoters. Together these results suggest that in particular formation of Smad2/3-Fra1 complexes may reflect activation of the Smad/AP-1-dependent TGFβ-induced invasion program.

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Acknowledgements

We thank our colleagues, in particular Aris Moustakas and Masato Morikawa, for their valuable discussion and help with experiments. This work was supported by the Swedish Cancerfonden (090773) and the Netherlands Centre for Biomedical Genetics.

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Author notes

  1. A Zieba and E Vasilaki: These authors contributed equally to this work.

Authors and Affiliations

  1. Ludwig Institute for Cancer Research, Science for Life Laboratory, Uppsala University, Uppsala, Sweden

    A Sundqvist, E Vasilaki, C Herrera Hidalgo, K Miyazono, C-H Heldin, P ten Dijke & H van Dam

  2. Department of Immunology, Genetics and Pathology, Science for Life Laboratory, Rudbeck Laboratory, Uppsala University, Uppsala, Sweden

    A Zieba, O Söderberg & U Landegren

  3. Department of Molecular Pathology, Graduate School of Medicine, University of Tokyo, Tokyo, Japan

    D Koinuma & K Miyazono

  4. Department of Molecular Cell Biology, Centre for Biomedical Genetics, Leiden University Medical Center, Leiden, The Netherlands

    P ten Dijke & H van Dam

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  1. A Sundqvist
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Correspondence to H van Dam.

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Competing interests

UL is a founder of Olink Bioscience, which commercializes the in situ PLA technology.

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Supplementary Information accompanies the paper on the Oncogene website

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Sundqvist, A., Zieba, A., Vasilaki, E. et al. Specific interactions between Smad proteins and AP-1 components determine TGFβ-induced breast cancer cell invasion. Oncogene 32, 3606–3615 (2013). https://doi.org/10.1038/onc.2012.370

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  • DOI: https://doi.org/10.1038/onc.2012.370

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