title: Cancer stem-like cells from head and neck cancers are chemosensitized by the Wnt antagonist, sFRP4, by inducing apoptosis, decreasing stemness, drug resistance and epithelial to mesenchymal transition.
abstract: Cancer stem cells (CSCs) of head and neck squamous cell carcinoma (HNSCC) are defined by high self-renewal and drug refractory potential. Involvement of Wnt/β-catenin signaling has been implicated in rapidly cycling cells such as CSCs, and inhibition of the Wnt/β-catenin pathway is a novel approach to target CSCs from HNSCC. In this study, we found that an antagonist of FrzB/Wnt, the secreted frizzled-related protein 4 (sFRP4), inhibited the growth of CSCs from two HNSCC cell lines, Hep2 and KB. We enriched the CD44(+) CSC population, and grew them in spheroid cultures. sFRP4 decreased the proliferation and increased the sensitivity of spheroids to a commonly used drug in HNSCC, namely cisplatin. Self-renewal in sphere formation assays decreased upon sFRP4 treatment, and the effect was reverted by the addition of Wnt3a. sFRP4 treatment of spheroids also decreased β-catenin, confirming its action through the Wnt/β-catenin signaling pathway. Quantitative PCR demonstrated a clear decrease of the stemness markers CD44 and ALDH, and an increase in CD24 and drug-resistance markers ABCG2 and ABCC4. Furthermore, we found that after sFRP4 treatment, there was a reversal in the expression of epithelial to mesenchymal (EMT) markers with the restoration of the epithelial marker E-cadherin, and depletion of EMT-specific markers twist, snail and N-cadherin. This is the first report demonstrating that the naturally occurring Wnt inhibitor, sFRP4, can be a potential drug to destroy CSC-enriched spheroids from HNSCCs. The repression of EMT and the decrease in stemness profile further strengthen the use of sFRP4 as a potent therapeutic against CSCs.
authors.affiliation: 1] Division of Cancer Stem Cells and Cardiovascular Regeneration, Manipal Institute of Regenerative Medicine, Manipal University, Bangalore, India [2] Faculty of Health Sciences, School of Biomedical Sciences, Curtin University, Perth, Western Australia.
authors.forename: S
authors.initials: S
authors.lastname: Warrier
authors.affiliation: Division of Cancer Stem Cells and Cardiovascular Regeneration, Manipal Institute of Regenerative Medicine, Manipal University, Bangalore, India.
authors.forename: G
authors.initials: G
authors.lastname: Bhuvanalakshmi
authors.affiliation: 1] Faculty of Health Sciences, School of Biomedical Sciences, Curtin University, Perth, Western Australia [2] School of Anatomy, Physiology and Human Biology, The University of Western Australia, Perth, Western Australia.
authors.forename: F
authors.initials: F
authors.lastname: Arfuso
authors.affiliation: School of Surgery, The University of Western Australia, Perth, Western Australia.
authors.forename: G
authors.initials: G
authors.lastname: Rajan
authors.affiliation: School of Medicine and Pharmacology, The University of Western Australia, Perth, Western Australia.
authors.forename: M
authors.initials: M
authors.lastname: Millward
authors.affiliation: Faculty of Health Sciences, School of Biomedical Sciences, Curtin University, Perth, Western Australia.
authors.forename: A
authors.initials: A
authors.lastname: Dharmarajan
mesh_terms: D000954:Antigens, Surface / Q000378:metabolism; D017209:Apoptosis / Q000187:drug effects*; D045744:Cell Line, Tumor; D019008:Drug Resistance, Neoplasm / Q000235:genetics; D058750:Epithelial-Mesenchymal Transition / Q000187:drug effects / Q000235:genetics; D006258:Head and Neck Neoplasms / Q000378:metabolism* / Q000473:pathology*; D006801:Humans; D016130:Immunophenotyping; D014411:Neoplastic Stem Cells / Q000187:drug effects* / Q000378:metabolism*; D011518:Proto-Oncogene Proteins / Q000494:pharmacology*; D018874:Spheroids, Cellular; D014407:Tumor Cells, Cultured; D051153:Wnt Proteins / Q000037:antagonists & inhibitors*; D060449:Wnt Signaling Pathway / Q000187:drug effects; D000094742:Secreted Frizzled-Related Proteins
chemical_list: D000954:Antigens, Surface; D011518:Proto-Oncogene Proteins; D051153:Wnt Proteins; C118723:SFRP4 protein, human; D000094742:Secreted Frizzled-Related Proteins
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