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Vimentin regulates EMT induction by Slug and oncogenic H-Ras and migration by governing Axl expression in breast cancer

Oncogene volume 30, pages 1436–1448 (2011)Cite this article

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Abstract

Epithelial-to-mesenchymal transition (EMT) is a critical event in the progression toward cancer metastasis. The intermediate filament protein vimentin is an important marker of EMT and a requisite regulator of mesenchymal cell migration. However, it is not known how vimentin functionally contributes to cancer cell invasion. Here, we report that ectopic expression of oncogenic H-Ras-V12G and Slug induces vimentin expression and migration in pre-malignant breast epithelial cells. Conversely, vimentin expression is necessary for Slug- or H-Ras-V12G-induced EMT-associated migration. Furthermore, silencing of vimentin in breast epithelial cells results in specific changes in invasiveness-related gene expression including upregulation of RAB25 (small GTPase Rab25) and downregulation of AXL (receptor tyrosine kinase Axl), PLAU (plasminogen activator, urokinase) and ITGB4 (integrin β4-subunit). Importantly, gene expression profiling analyses reveal that vimentin expression correlates positively/negatively with these genes also in multiple breast cancer cell lines and breast cancer patient samples. Focusing on the tyrosine kinase Axl, we show that induction of vimentin by EMT is associated with upregulation of Axl expression and that Axl enhances the migratory activity of pre-malignant breast epithelial cells. Using null and knock-down cells and overexpression models, we also show that regulation of breast cancer cell migration in two- and three-dimensional matrices by vimentin is Axl- dependent and that Axl functionally contributes to lung extravasation of breast cancer cells in mice. In conclusion, our data show that vimentin functionally contributes to EMT and is required for induction of Axl expression. Moreover, these results provide a molecular explanation for vimentin-dependent cancer cell migration during EMT by identifying Axl as a key proximal component in this process.

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Acknowledgements

We thank H Marttila, J Siivonen, L Lahtinen, J Rantala, P Terho, E Mattila and T Hoiby for their excellent technical assistance, H Sara for the bioinformatic analysis of the cDNA microarray, K Hellström, S-K Leivonen and the Finnish DNA Microarray Centre for their help with gene expression analysis and TCDM (Turku) for the animal experiments. M Salmi is acknowledged for the Hb116 antibody and for critically reading and editing the manuscript. This work was supported by grants from the Academy of Finland, the Sigrid Juselius Foundation, Emil Aaltonen Foundation, Finnish Cancer Organisations, EMBO Young Investigator Program, University of Turku, Turku University Foundation, Finnish Concordia Fund, Finnish Cultural Foundation, Ida Montini Foundation, Orion and Farmos Research Foundation, K Albin Johansson's Foundation and Scientific Foundation of Instrumentarium. K Vuoriluoto is financially supported by Turku Graduate School for Biomedical Sciences.

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Authors and Affiliations

  1. Medical Biotechnology, VTT Technical Research Centre of Finland, Turku, Finland

    K Vuoriluoto, S Kiviluoto, J-P Mpindi, J Nevo & J Ivaska

  2. Turku Centre for Biotechnology, University of Turku, Turku, Finland

    K Vuoriluoto, J Nevo & J Ivaska

  3. Department of Biomedicine, University of Bergen, Bergen, Norway

    H Haugen, C Gjerdrum, C Tiron & J B Lorens

  4. Department of Biochemistry and Food Chemistry, University of Turku, Turku, Finland

    J Ivaska

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  1. K Vuoriluoto
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Correspondence to J Ivaska.

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Vuoriluoto, K., Haugen, H., Kiviluoto, S. et al. Vimentin regulates EMT induction by Slug and oncogenic H-Ras and migration by governing Axl expression in breast cancer. Oncogene 30, 1436–1448 (2011). https://doi.org/10.1038/onc.2010.509

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