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Salinomycin induces cell death and differentiation in head and neck squamous cell carcinoma stem cells despite activation of epithelial-mesenchymal transition and Akt

Selena Z Kuo1, Katherine J Blair1, Elham Rahimy1, Alan Kiang1, Eric Abhold1, Jian-Bing Fan2, Jessica Wang-Rodriguez3, Xabier Altuna4 and Weg M Ongkeko1*

Author Affiliations

1 Division of Otolaryngology-Head and Neck Surgery, Department of Surgery, University of California, San Diego, San Diego, CA, USA

2 Illumina Inc., San Diego, CA, 92121, USA

3 Veterans Administration Medical Center and Department of Pathology, University of California, San Diego, La Jolla, CA, USA

4 Hospital Universitario Donostia, San Sebastian, Spain

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BMC Cancer 2012, 12:556  doi:10.1186/1471-2407-12-556

Published: 24 November 2012

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Additional File 1:

Format: PDF. Cell death does not significantly alter expression of EMT and stem cell genes in JLO-1. A drug control was used to confirm that dose-dependent induction of EMT genes and repression of stem cell genes was not a mere epiphenomenon of cell death accompanying salinomycin treatment. We have previously discovered that Metformin does not influence EMT in JLO-1 cells at non-cytotoxic doses, indicating it does not regulate EMT in JLO-1 (unpublished data). (A) An MTS assay was initially performed to determine the cytoxicity curve for JLO-1 cells treated with Metformin for 72 hours. (B) At non–cytotoxic concentrations, Metformin does not regulate EMT based on RT-qPCR data of Snail and E-cadherin transcript levels, thus it is an appropriate drug control to induce cell death in JLO-1. (C) Upon 48-hour treatment of JLO-1 with 15 mM Metformin to induce approximately 60% cell death (equivalent to the cell death observed from 4 μM salinomycin treatment), expression of EMT genes Snail and E-cadherin showed only minor changes. In addition, CD44 expression was not effected by induction of cell death.

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