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  • Original Article
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Loss of breast epithelial marker hCLCA2 promotes epithelial-to-mesenchymal transition and indicates higher risk of metastasis

Oncogene volume 31, pages 2237–2246 (2012)Cite this article

Abstract

Transition between epithelial and mesenchymal states is a feature of both normal development and tumor progression. We report that expression of chloride channel accessory protein hCLCA2 is a characteristic of epithelial differentiation in the immortalized MCF10A and HMLE models, while induction of epithelial-to-mesenchymal transition by cell dilution, TGFβ or mesenchymal transcription factors sharply reduces hCLCA2 levels. Attenuation of hCLCA2 expression by lentiviral small hairpin RNA caused cell overgrowth and focus formation, enhanced migration and invasion, and increased mammosphere formation in methylcellulose. These changes were accompanied by downregulation of E-cadherin and upregulation of mesenchymal markers such as vimentin and fibronectin. Moreover, hCLCA2 expression is greatly downregulated in breast cancer cells with a mesenchymal or claudin-low profile. These observations suggest that loss of hCLCA2 may promote metastasis. We find that higher-than-median expression of hCLCA2 is associated with a one-third lower rate of metastasis over an 18-year period among breast cancer patients compared with lower-than-median (n=344, unfiltered for subtype). Thus, hCLCA2 is required for epithelial differentiation, and its loss during tumor progression contributes to metastasis. Overexpression of hCLCA2 has been reported to inhibit cell proliferation and is accompanied by increases in chloride current at the plasma membrane and reduced intracellular pH (pHi). We found that knockdown cells have sharply reduced chloride current and higher pHi, both characteristics of tumor cells. These results suggest a mechanism for the effects on differentiation. Loss of hCLCA2 may allow escape from pHi homeostatic mechanisms, permitting the higher intracellular and lower extracellular pH that are characteristic of aggressive tumor cells.

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Acknowledgements

This work was supported by the following grants: NIH grant 1R15CA151094-01 and Excellence in Academic Medicine grants 56252 and 56253 to RCE; and NIH grants DK065742 and DA028017 to LP.

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Author notes

  1. V Walia

    Present address: 7Current address: Cancer Genetics Branch, National Human Genome Research Institute, National Institutes of Health (NIH), Bethesda, MD, USA.,

Authors and Affiliations

  1. Department of Pharmacology, Southern Illinois University School of Medicine, Springfield, IL, USA

    V Walia, D Cao, L Premkumar & R C Elble

  2. Simmons Cancer Institute, Southern Illinois University School of Medicine, Springfield, IL, USA

    V Walia, Y Yu, J Cheng, K Rao & R C Elble

  3. Committee on Genetics, Genomics and Systems Biology (GGSB), University of Chicago, Chicago, IL, USA

    M Sun

  4. Department of Cell and Developmental Biology, Vanderbilt University School of Medicine, Nashville, TN, USA

    J R McLean

  5. Institute of Health and Biomedical Innovation, Queensland University of Technology, Brisbane, Queensland, Australia

    B G Hollier

  6. Department of Molecular Pathology, The University of Texas M.D. Anderson Cancer Center, Houston, TX, USA

    S A Mani

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Correspondence to R C Elble.

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Walia, V., Yu, Y., Cao, D. et al. Loss of breast epithelial marker hCLCA2 promotes epithelial-to-mesenchymal transition and indicates higher risk of metastasis. Oncogene 31, 2237–2246 (2012). https://doi.org/10.1038/onc.2011.392

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