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α3β1 integrins regulate CD151 complex assembly and membrane dynamics in carcinoma cells within 3D environments

Oncogene volume 32, pages 3965–3979 (2013)Cite this article

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Abstract

Integrins are extracellular matrix (ECM) receptors that are key players in the regulation of tumour cell invasion. The laminin-binding integrin α3β1 has previously been shown to regulate adhesion and migration of carcinoma cells in part through co-operative signalling with the tetraspanin family of transmembrane proteins. However, the spatial and temporal regulation of crosstalk between these families of transmembrane proteins in intact cells remains poorly understood. Here we have used fluorescence resonance energy transfer (FRET) to demonstrate for the first time that α3β1 and the tetraspanin CD151 directly associate at the front and retracting rear of polarised migrating breast carcinoma cells in both two-dimentional (2D) and three-dimentional (3D)matrices. Furthermore, localised α3β1–CD151 binding correlates with lower CD151 homodimerisation in cells migrating on laminin or within matrigel. Loss of α3β1 integrin leads to increased CD151 homodimer formation, increased activation of Rho GTPase, loss of cell polarity and decreased invasion in 3D ECM. As a result, α3-silenced cells show decreased actin-based membrane protrusion and retraction in both 2D and 3D environments. These data demonstrate that associations between α3β1 and CD151 occur dynamically within discrete subcellular compartments and act to establish local GTPase signalling to promote tumour cell invasion. These novel findings shed light on the complex crosstalk and switching between receptor complexes in response to different extracellular cues during cell invasion in 3D environments.

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Acknowledgements

We would like to thank Simon Ameer-Beg and Daniel Matthews for technical assistance and support with the multiphoton FLIM. This research was funded by grants from Cancer Research UK (to MP, FB and NH), the Wellcome Trust (to NH), the Biotechnology and Biological Sciences Research Council and Engineering and Physical Sciences Research Council (to BO) and a Royal Society University Research Fellowship (to MP).

Author contributions: TMES and MP designed and performed the experiments and wrote the manuscript. BO and MRH wrote software and analysed the protrusion and retraction behaviour in 3D and 2D, respectively. FB generated a number of the expression plasmids. NAH and FB participated in the design of the study and assisted in drafting the manuscript. All authors read and approved the final manuscript.

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Authors and Affiliations

  1. Randall Division of Cell and Molecular Biophysics, King’s College London, New Hunts House, Guys Campus, London, UK

    T M E Scales, A Jayo, M R Holt & M Parsons

  2. Oxford e-Research Centre and Oxford Centre for Integrative Systems Biology, University of Oxford, Oxford, UK

    B Obara

  3. Cancer Research UK Institute for Cancer Studies, University of Birmingham, Birmingham, UK

    N A Hotchin

  4. School of Biosciences, University of Birmingham, Birmingham, UK

    F Berditchevski

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  1. T M E Scales
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Correspondence to M Parsons.

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Scales, T., Jayo, A., Obara, B. et al. α3β1 integrins regulate CD151 complex assembly and membrane dynamics in carcinoma cells within 3D environments. Oncogene 32, 3965–3979 (2013). https://doi.org/10.1038/onc.2012.415

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