Maintenance of stem cell self-renewal in head and neck cancers requires actions of GSK3β influenced by CD44 and RHAMM

Authors

  • Hideo Shigeishi,

    1. Blizard Institute Barts and the London School of Medicine and Dentistry, Queen Mary University of London, London United Kingdom
    2. Department of Oral and Maxillofacial Surgery, Division of Cervico-Gnathostomatology, Graduate School of Biomedical Sciences, Hiroshima University, Hiroshima, Japan
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  • Adrian Biddle,

    1. Blizard Institute Barts and the London School of Medicine and Dentistry, Queen Mary University of London, London United Kingdom
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  • Luke Gammon,

    1. Blizard Institute Barts and the London School of Medicine and Dentistry, Queen Mary University of London, London United Kingdom
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  • Helena Emich,

    1. Blizard Institute Barts and the London School of Medicine and Dentistry, Queen Mary University of London, London United Kingdom
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  • Camila O. Rodini,

    1. Blizard Institute Barts and the London School of Medicine and Dentistry, Queen Mary University of London, London United Kingdom
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  • Emilios Gemenetzidis,

    1. Blizard Institute Barts and the London School of Medicine and Dentistry, Queen Mary University of London, London United Kingdom
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  • Bilal Fazil,

    1. Blizard Institute Barts and the London School of Medicine and Dentistry, Queen Mary University of London, London United Kingdom
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  • Masaru Sugiyama,

    1. Department of Public Oral Health, Division of Oral Health Science, Graduate School of Biomedical Sciences, Hiroshima University, Hiroshima, Japan
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  • Nobuyuki Kamata,

    1. Department of Oral and Maxillofacial Surgery, Division of Cervico-Gnathostomatology, Graduate School of Biomedical Sciences, Hiroshima University, Hiroshima, Japan
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  • Ian C. Mackenzie

    Corresponding author
    1. Blizard Institute Barts and the London School of Medicine and Dentistry, Queen Mary University of London, London United Kingdom
    • BDS, FDSRCS, Ph.D., Blizard Institute, Barts and The London School of Medicine and Dentistry, Queen Mary University of London, 4 Newark Street, London E1 2AT, U.K. E-mail: i.c.mackenzie@qmul.ac.uk Telephone: 20-7882-7159; Fax: 20-7882-7172

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  • Author contributions: H.S.: conception and design, financial support, administrative support, collection and/or assembly of data, data analysis and interpretation, manuscript writing, and final approval of manuscript; A.B. and L.G.: conception and design, data analysis and interpretation, manuscript writing, and final approval of manuscript; H.E., C.O.R., E.G., and B.F.: data analysis and interpretation and final approval of manuscript; M.S. and N.K.: conception and design, financial support, administrative support, and final approval of manuscript; I.C.M.: conception and design, financial support, data analysis and interpretation, manuscript writing, and final approval of manuscript.

Abstract

Cells sorted from head and neck cancers on the basis of their high expression of CD44 have high potency for tumor initiation. These cells are also involved in epithelial to mesenchymal transition (EMT) and we have previously reported that cancer stem cells (CSCs) exist as two biologically distinct phenotypes. Both phenotypes are CD44high but one is also ESAhigh and maintains epithelial characteristics, the other is ESAlow, has mesenchymal characteristics and is migratory. Examining CD44-regulated signal pathways in these cells we show that CD44, and also RHAMM, act to inhibit phosphorylation of glycogen synthase kinase 3β (GSK3β). We show that inhibitory phosphorylation reduces the formation of both “tumor spheres” and “holoclone” colonies, functional indicators of stemness. GSK3β inhibition also reduces the expression of stem cell markers such as Oct4, Sox2, and Nanog and upregulates expression of the differentiation markers Calgranulin B and Involucrin in the CD44high/ESAhigh cell fraction. Transition of CSCs out of EMT and back to the epithelial CSC phenotype is induced by GSK3β knockdown. These results indicate that GSK3β plays a central role in determining and maintaining the phenotypes and behavior of CSCs in vitro and are likely to be involved in controlling the growth and spread of tumors in vivo.

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