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Loss of Par3 promotes breast cancer metastasis by compromising cell–cell cohesion

Nature Cell Biology volume 15, pages 189–200 (2013)Cite this article

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Abstract

The mechanisms by which tumour cells metastasize and the role that cell polarity proteins play in this process are not well understood. We report that partitioning defective protein 3 (Par3) is dysregulated in metastasis in human breast cancer, and is associated with a higher tumour grade and ErbB2-positive status. Downregulation of Par3 cooperated with ErbB2 to induce cell invasion and metastasis in vivo. Interestingly, the metastatic behaviour was not associated with an overt mesenchymal phenotype. However, loss of Par3 inhibited E-cadherin junction stability, disrupted membrane and actin dynamics at cell–cell junctions and decreased cell–cell cohesion in a manner dependent on the Tiam1/Rac–GTP pathway. Inhibition of this pathway restored E-cadherin junction stability and blocked invasive behaviour of cells lacking Par3, suggesting that loss of Par3 promotes metastatic behaviour of ErbB2-induced tumour epithelial cells by decreasing cell–cell cohesion.

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Figure 1: Loss of Par3 cooperates with ErbB2 to induce invasive behaviour in mammary epithelial cells.
Figure 2: Downregulation of Par3 promotes metastasis without EMT.
Figure 3: Loss of Par3 weakens cell–cell adhesions and inhibits E-cadherin junction stability.
Figure 4: Loss of Par3 induces aberrant activation of Tiam1–Rac signalling.
Figure 5: Contribution of Tiam1–Rac and ErbB2 signalling to the Par3-loss-induced phenotype.
Figure 6: Loss of Par3 induces changes in actin organization and mislocalizes the Arp2/3 complex.
Figure 7: Loss of Par3 introduces changes in actin and E-cadherin dynamics at cell–cell junctions.
Figure 8: Dysregulation of Par3 in human breast cancer.

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Acknowledgements

The authors thank W. J. Muller (McGill University, Canada) for the NDL transgenic mice, I. Macara (Univ of Virginia, USA) for the Tiam1 cDNA, M. Matsuda (Kyoto University, Japan) for the Raichu-Rac plasmid and D. Barber (UCSF, USA) for sharing the anti-ARPC2 antibody. We thank J. Haynes for critical reading of the manuscript and M. Egeblad and R. Sordella for helpful comments. We thank S. Hearn for assistance with microscopy and C. Camarda for assistance with the graphic abstract. We thank members of the Muthuswamy laboratory for critical comments on the manuscripts and insightful discussions. This work was supported by CA098830, BC075024, Era of Hope Scholar award from DOD Breast Cancer Research Program; Rita Allen Foundation, Lee K Margaret Lau Chair for breast cancer research and Campbell Family Institute for Breast cancer research to S.K.M. This was also financially supported in part by the Ontario Ministry of Health and Long Term Care. The views expressed do not necessarily reflect those of the OMOHLTC.

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Authors and Affiliations

  1. Cold Spring Harbor Laboratory, One Bungtown Road, Cold Spring Harbor, New York 11724, USA

    Bin Xue, Kannan Krishnamurthy & Senthil K. Muthuswamy

  2. Department of Molecular and Cellular Biology, Stony Brook University, Stony Brook, New York 11974, USA

    Bin Xue & Senthil K. Muthuswamy

  3. Department of Pathology and Immunology, Washington University School of Medicine, St Louis, Missouri 63110, USA

    D. Craig Allred

  4. Ontario Cancer Institute, Campbell Family Institute for Breast Cancer Research, University of Toronto, Room 9-303, 610 University Avenue, Toronto, Ontario M5G 2M9, Canada

    Senthil K. Muthuswamy

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Contributions

K.K. performed Transwell invasion assays in Figs 1d,g 5g, B.X. conducted the rest of the experiments. S.K.M. conceived and directed the study, B.X. and S.K.M. prepared the manuscript. D.C.A. provided the human tissues, read, quantified and interpreted the IHC analysis.

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Correspondence to Senthil K. Muthuswamy.

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Actin and E-cadherin dynamics at cell-cell junctions.

10A. control or 10A. Par3-shRNA cells expressing E-cad–GFP and LifeAct–RFP were imaged in the middle of the cell (1.5 mm above the bottom) at the rate of 12 frames per minute for 10 minutes. (MOV 2913 kb)

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Xue, B., Krishnamurthy, K., Allred, D. et al. Loss of Par3 promotes breast cancer metastasis by compromising cell–cell cohesion. Nat Cell Biol 15, 189–200 (2013). https://doi.org/10.1038/ncb2663

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