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. 2017 Jul;14(1):1147-1151.
doi: 10.3892/ol.2017.6189. Epub 2017 May 17.

Effect of thymoquinone on head and neck squamous cell carcinoma cells in vitro: Synergism with radiation

Ulana Kotowski  1 , Gregor Heiduschka  1 , Lorenz Kadletz  1 , Tammer Fahim  1 , Rudolf Seemann  2 , Rainer Schmid  3 , Sven Schneider  1 , Andreas Mitterbauer  4 , Dietmar Thurnher  5
Affiliations

Effect of thymoquinone on head and neck squamous cell carcinoma cells in vitro: Synergism with radiation

Ulana Kotowski et al. Oncol Lett. 2017 Jul.

Abstract

Thymoquinone (TQ) is the main bioactive constituent present in black seed oil (Nigella sativa); it has shown anti-inflammatory and anti-neoplastic effects in various cancer cell types. The aim of the present study was to investigate the effects of TQ on head and neck squamous cell carcinoma (HNSCC) cell lines, on its own and in combination with radiation and cisplatin, respectively. The SCC25 and CAL27 HNSCC cell lines were treated with TQ alone and in combination with cisplatin or radiation, respectively. Proliferation assays and clonogenic assays were performed. Apoptosis was detected by flow cytometry. TQ exhibited dose-dependent cytotoxicity via apoptosis in the investigated cell lines. In combination with cisplatin, TQ resulted in no significant increase in cytotoxicity. Combined with radiation, TQ significantly reduced clonogenic survival compared with each treatment method alone. TQ is a promising agent in the treatment of head and neck cancer due to its anti-proliferative and radiosensitizing properties. However, the combination of TQ with cisplatin showed no therapeutic benefit in vitro.

Keywords: cisplatin; head and neck squamous cell carcinoma; radiation; radiosensitizer; thymoquinone.

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Figures

Figure 1.
Figure 1.
Dose-response curves of SCC25 and CAL27 cells following treatment with increasing doses of thymoquinone (TQ) and 72 h of incubation.
Figure 2.
Figure 2.
Combination index (CI) plot for the combination of thymoquinone and cisplatin. CI values where calculated by CalcuSyn® software. CI=1 indicates an additive effect, CI<1 indicates a synergistic effect and CI>1 indicates antagonism.
Figure 3.
Figure 3.
Clonogenic survival of cells treated with thymoquinone (TQ) alone and in combination with radiation. The cells were treated with TQ (SCC25 with 0, 10 or 20 µM; and CAL27 with 0, 5 or 10 µM) and then irradiated with 0, 2, 4, 6 or 8 Gy. Surviving colonies were counted after 10 days and expressed as the surviving fraction.
Figure 4.
Figure 4.
Evaluation of apoptosis by flow cytometry. The cells were treated with thymoquinone (TQ) (SCC25 with 0, 40 and 60 µM; and CAL27 with 0, 20 and 40 µM). Apoptosis was measured after 48 h by flow cytometry using the Annexin-V Apoptosis Detection kit. *P=0.0021 for SCC25 cells treated with 40 µM TQ + 2 Gy vs. SCC25 cells treated with 2 Gy only, P=0.0640 for CAL27 cells treated with 10 µM TQ vs. control cells and P<0.0001 for CAL27 cells treated with 20 µM TQ vs. control cells.
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