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    Investigation of the effects of 5-fluorouracil and thymoquinone on head and neck cancer stem cells
    (Canadian Soc Clinical Investigation, 2015) Acar, Muradiye; Gunduz, Mehmet; Ocak, Tarik; Gurses, Hacer Esra; Cetin, Elif Nihan; Ceylan, Furkan Sacit; Sefa, Alperen
    Purpose: A subpopulation of cancer stem cells (CSCs) in head and neck squamous cell carcinoma (HNSCC) play a critical role in invasion, metastasis, and tumour recurrence post-therapy. 5-Fluorouracil (5-FU) is a pyrimidine analogue that is used in the treatment of HNSCC. Thymoquinone (TQ), found in Cumin (Nigella sativa) seed oil, has anti-diabetic, anti-oxidant, anti-inflammatory, and antitumour effects. The purpose of this study was to examine the effects of 5-FU and TQ on CSCs. Methods: ALDH1 (+) cells were isolated from the HNSCC cell line UT-SCC-74A using magnetic activated cell sorting (MACS) and sphere formation was confirmed. Real-time RTPCR revealed was used to measure expression levels of a number of stem cell markers, including OCT4, SOX2, and KLF-4. To investigate the effects of the therapeutic agents, 5-FU and TQ, concentration-dependent assay procedures were used and cell viability after treatment was measured using XTT assay. Results: ALDH1 (+) cells formed spheres while the ALDH1 (-) cells did not. Real-time RT-PCR revealed that a number of stem cell markers, including OCT4, SOX2, and KLF-4, were up-regulated in the CSCs. XTT results showed that, when given alone at a dose of 50 ng/mL, neither agent had a statistically significant cytotoxic effect on CSCs. However, when dosed in combination, TQ and 5-FU had a synergistic effect on CSC cytotoxicity. Conclusion: We report the effects of TQ and 5-FU on CSCs in HNSCC.