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Mitochondrial SOD2 regulates epithelial–mesenchymal transition and cell populations defined by differential CD44 expression

Oncogene volume 34, pages 5229–5239 (2015)Cite this article

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Abstract

Epithelial–mesenchymal transition (EMT) promotes cancer cell invasion, metastasis and treatment failure. EMT may be activated in cancer cells by reactive oxygen species (ROS). EMT may promote conversion of a subset of cancer cells from a CD44low-CD24high (CD44L) epithelial phenotype to a CD44high-CD24−/low (CD44H) mesenchymal phenotype, the latter associated with increased malignant properties of cancer cells. ROS are required for cells undergoing EMT, although excessive ROS may induce cell death or senescence; however, little is known as to how cellular antioxidant capabilities may be regulated during EMT. Mitochondrial superoxide dismutase 2 (SOD2) is frequently overexpressed in oral and esophageal cancers. Here, we investigate mechanisms of SOD2 transcriptional regulation in EMT, as well as the functional role of this antioxidant in EMT. Using well-characterized genetically engineered oral and esophageal human epithelial cell lines coupled with RNA interference and flow cytometric approaches, we find that transforming growth factor (TGF)-β stimulates EMT, resulting in conversion of CD44L to CD44H cells, the latter of which display SOD2 upregulation. SOD2 induction in transformed keratinocytes was concurrent with suppression of TGF-β-mediated induction of both ROS and senescence. SOD2 gene expression appeared to be transcriptionally regulated by NF-κB and ZEB2, but not ZEB1. Moreover, SOD2-mediated antioxidant activity may restrict conversion of CD44L cells to CD44H cells at the early stages of EMT. These data provide novel mechanistic insights into the dynamic expression of SOD2 during EMT. In addition, we delineate a functional role for SOD2 in EMT via the influence of this antioxidant upon distinct CD44L and CD44H subsets of cancer cells that have been implicated in oral and esophageal tumor biology.

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Acknowledgements

We are grateful to the Molecular Pathology & Imaging, Molecular Biology/Gene Expression and Cell Culture Core Facilities of the NIH/NIDDK Center for Molecular Studies in Digestive and Liver Diseases (P30-DK050306) and of the NIH P01CA098101. This study was supported in part by NIH Grants P01CA098101 (to HK, KAW, KT, MN, AL, AG, SC, SK, JAD and HN), F32CA174176 (KAW), F30CA175133 (AL), K26RR032714 (HN), R01CA073599 (DKS) as well as Pennsylvania CURE Program Grant (HN), the Edward P. Evans Foundation (DKS), University of Pennsylvania University Research Foundation Award (HN) and University of Pennsylvania, Abramson Cancer Center Pilot Project Grant (HN).

Abbreviations

CSC, cancer stem cell; EMT, epithelial–mesenchymal transition; FACS, fluorescence-activated cell sorting; RNAi, RNA interference; ROS, reactive oxygen species; SOD2, superoxide dismutase 2; TGF-β, transforming growth factor-β

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Author notes

  1. H Kinugasa and K A Whelan: These authors contributed equally to this work.

Authors and Affiliations

  1. Gastroenterology Division, Department of Medicine, University of Pennsylvania, Philadelphia, PA, USA

    H Kinugasa, K A Whelan, K Tanaka, M Natsuizaka, A Long, A Guo, S Chang, S Kagawa & H Nakagawa

  2. Abramson Cancer Center, University of Pennsylvania, Philadelphia, PA, USA

    H Kinugasa, K A Whelan, K Tanaka, M Natsuizaka, A Long, A Guo, S Chang, S Kagawa, J A Diehl & H Nakagawa

  3. Department of Gastroenterology and Hepatology, Okayama University Graduate School of Medicine, Dentistry, and Pharmaceutical Sciences, Okayama, Japan

    H Kinugasa & K Yamamoto

  4. Department of Gastroenterology and Hepatology, Hokkaido University, Sapporo, Japan

    M Natsuizaka

  5. Department of Animal Biology and Mari Lowe Center for Comparative Oncology, School of Veterinary Medicine, University of Pennsylvania, Philadelphia, PA, USA

    S Srinivasan, M Guha & N G Avadhani

  6. Department of Toxicology and Cancer Biology, University of Kentucky College of Medicine, Lexington, KY, USA

    D K St Clair

  7. Department of Biochemistry and Molecular Biology, Medical University of South Carolina, Charleston, SC, USA

    J A Diehl

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  1. H Kinugasa
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Correspondence to H Nakagawa.

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Kinugasa, H., Whelan, K., Tanaka, K. et al. Mitochondrial SOD2 regulates epithelial–mesenchymal transition and cell populations defined by differential CD44 expression. Oncogene 34, 5229–5239 (2015). https://doi.org/10.1038/onc.2014.449

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