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Cytotoxic, apoptotic and genotoxic effects of thymoquinone‑oxime derivative on gastric cancer cells: An in vitro study

  • Authors:
    • Kübra Bozali̇
    • Zeynep İnce
    • Nurhi̇lal Kiziltoprak
    • Taygun Gülşen
    • Mehmet Köstek
    • Yasi̇r Musa Kesgi̇n
    • Muhammer Ergenç
    • Mi̇ne Dağgez
    • Eren Altun
    • Fati̇h Taşkesen
    • Cebrai̇l Akyüz
    • Oğuzhan Sunamak
    • Mustafa Duman
    • Eray Meti̇n Güler
  • View Affiliations / Copyright

    Affiliations: Department of Medical Biochemistry, Faculty of Hamidiye Medicine, University of Health Sciences, Istanbul 34668, Türkiye, Department of General Surgery, University of Health Sciences, Sultan 2. Abdulhamid Han Training and Research Hospital, Istanbul 34668, Türkiye, Department of Molecular Oncology, Hamidiye Institute of Health Sciences, University of Health Sciences, Istanbul 34668, Türkiye, Department of General Surgery, Haydarpasa Numune Health Application and Research Center, Istanbul 34668, Türkiye
    Copyright: © Bozali̇ et al. This is an open access article distributed under the terms of Creative Commons Attribution License.
  • Article Number: 109
    |
    Published online on: January 14, 2026
       https://doi.org/10.3892/ol.2026.15462
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Abstract

Gastric cancer (GC) remains a notable global health concern, emphasizing the need for novel and effective therapeutic agents. The present study investigated the cytotoxic, genotoxic and apoptotic effects of thymoquinone‑oxime (TQ‑ox) on human gastric adenocarcinoma AGS cells and assessed its potential to induce DNA damage. Cytotoxicity was evaluated in AGS and normal human gastric epithelial cells (HGEpiCs) using a luminometric ATP assay. Intracellular reactive oxygen species (ROS) and mitochondrial membrane potential (MMP) were measured fluorometrically, whereas intracellular glutathione (GSH) content was determined via a luminometric GSH assay kit. DNA damage was quantified by a comet assay, and apoptosis was assessed by fluorescence microscopy with acridine orange/ethidium bromide (AO/EB) double staining. TQ‑ox exhibited dose‑dependent cytotoxicity in gastric cells, with AGS cells having a slightly higher sensitivity to TQ‑ox treatment than HGEpiCs [half‑maximal inhibitory concentration (IC50): 40.29 vs. 46.42 µM]. Treatment with TQ‑ox significantly increased intracellular ROS levels and DNA damage, while also inducing a significant depletion of intracellular GSH and a reduction of MMP, indicating an increase in oxidative stress (OS) and mitochondrial dysfunction. AO/EB staining supported a dose‑dependent increase in apoptosis of gastric cells at sub‑IC50 concentrations of TQ‑ox. Similarly, comet assay results revealed greater genotoxic effects in AGS cells compared with HGEpiCs, particularly at higher doses of TQ‑ox. These findings demonstrated that TQ‑ox exerted cytotoxic, pro‑apoptotic and genotoxic effects on GC cells, likely mediated by OS, mitochondrial impairment and DNA damage. Taken together, these results provide additional evidence supporting the mechanistic effects of TQ‑ox on GC cells and highlighted its potential as a candidate molecule for further preclinical evaluation.

View Figures

Figure 1

(A) Cell viability decreases as the
dose increases. (B) Intracellular ROS levels increase significantly
as the thymoquinone-oxime concentration increases in both cell
lines. Statistical significance was analyzed using one-way analysis
of variance with Tukey's post hoc test. *P<0.05, **P<0.01 and
***P<0.001 vs. control. RLU, relative luminescence units; ROS,
reactive oxygen species; RFU, relative fluorescence units; HGEpiC,
human gastric epithelial cell.

Figure 2

Effects of thymoquinone-oxime
treatment on (A) GSH (µM) levels and (B) MMP (ΔΨm). Statistical
significance was analyzed using one-way analysis of variance with
Tukey's post hoc test. *P<0.05, **P<0.01 and ***P<0.001
vs. control. HGEpiC, human gastric epithelial cell; GSH,
glutathione; MMP, mitochondrial membrane potential.

Figure 3

Effect of TQ-ox on apoptosis in
HGEpiCs and AGS cells. Statistical significance was analyzed using
one-way analysis of variance with Tukey's post hoc test.
**P<0.01 and ***P<0.001 vs. control. Representative
fluorescence microscopy images of acridine orange/ethidium
bromide-stained cells showing live (green) and apoptotic
(red/orange) cells in the control and 40 µM TQ-ox treated AGS
groups. A total of ~50 cells were analyzed per condition. Scale
bar, 100 µm. HGEpiC, human gastric epithelial cell; TQ-ox,
thymoquinone-oxime.

Figure 4

Effect of TQ-ox on DNA damage in
HGEpiCs and AGS cells following 24 h of TQ-ox treatment (5–40 µM).
Statistical significance was analyzed using one-way analysis of
variance with Tukey's post hoc test. **P<0.01 and ***P<0.001
vs. control. Representative fluorescence microscopy images showing
nuclei (control) and comet tails (40 µM TQ-ox-treated cells),
indicating fragmented DNA. A total of ~50 cells were analyzed per
condition. Scale bar, 200 µm. HGEpiC, human gastric epithelial
cell; TQ-ox, thymoquinone-oxime.
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Copy and paste a formatted citation
Spandidos Publications style
Bozali̇ K, İnce Z, Kiziltoprak N, Gülşen T, Köstek M, Kesgi̇n YM, Ergenç M, Dağgez M, Altun E, Taşkesen F, Taşkesen F, et al: <p>Cytotoxic, apoptotic and genotoxic effects of thymoquinone‑oxime derivative on gastric cancer cells: An <em>in vitro</em> study</p>. Oncol Lett 31: 109, 2026.
APA
Bozali̇, K., İnce, Z., Kiziltoprak, N., Gülşen, T., Köstek, M., Kesgi̇n, Y.M. ... Güler, E.M. (2026). <p>Cytotoxic, apoptotic and genotoxic effects of thymoquinone‑oxime derivative on gastric cancer cells: An <em>in vitro</em> study</p>. Oncology Letters, 31, 109. https://doi.org/10.3892/ol.2026.15462
MLA
Bozali̇, K., İnce, Z., Kiziltoprak, N., Gülşen, T., Köstek, M., Kesgi̇n, Y. M., Ergenç, M., Dağgez, M., Altun, E., Taşkesen, F., Akyüz, C., Sunamak, O., Duman, M., Güler, E. M."<p>Cytotoxic, apoptotic and genotoxic effects of thymoquinone‑oxime derivative on gastric cancer cells: An <em>in vitro</em> study</p>". Oncology Letters 31.3 (2026): 109.
Chicago
Bozali̇, K., İnce, Z., Kiziltoprak, N., Gülşen, T., Köstek, M., Kesgi̇n, Y. M., Ergenç, M., Dağgez, M., Altun, E., Taşkesen, F., Akyüz, C., Sunamak, O., Duman, M., Güler, E. M."<p>Cytotoxic, apoptotic and genotoxic effects of thymoquinone‑oxime derivative on gastric cancer cells: An <em>in vitro</em> study</p>". Oncology Letters 31, no. 3 (2026): 109. https://doi.org/10.3892/ol.2026.15462
Copy and paste a formatted citation
x
Spandidos Publications style
Bozali̇ K, İnce Z, Kiziltoprak N, Gülşen T, Köstek M, Kesgi̇n YM, Ergenç M, Dağgez M, Altun E, Taşkesen F, Taşkesen F, et al: <p>Cytotoxic, apoptotic and genotoxic effects of thymoquinone‑oxime derivative on gastric cancer cells: An <em>in vitro</em> study</p>. Oncol Lett 31: 109, 2026.
APA
Bozali̇, K., İnce, Z., Kiziltoprak, N., Gülşen, T., Köstek, M., Kesgi̇n, Y.M. ... Güler, E.M. (2026). <p>Cytotoxic, apoptotic and genotoxic effects of thymoquinone‑oxime derivative on gastric cancer cells: An <em>in vitro</em> study</p>. Oncology Letters, 31, 109. https://doi.org/10.3892/ol.2026.15462
MLA
Bozali̇, K., İnce, Z., Kiziltoprak, N., Gülşen, T., Köstek, M., Kesgi̇n, Y. M., Ergenç, M., Dağgez, M., Altun, E., Taşkesen, F., Akyüz, C., Sunamak, O., Duman, M., Güler, E. M."<p>Cytotoxic, apoptotic and genotoxic effects of thymoquinone‑oxime derivative on gastric cancer cells: An <em>in vitro</em> study</p>". Oncology Letters 31.3 (2026): 109.
Chicago
Bozali̇, K., İnce, Z., Kiziltoprak, N., Gülşen, T., Köstek, M., Kesgi̇n, Y. M., Ergenç, M., Dağgez, M., Altun, E., Taşkesen, F., Akyüz, C., Sunamak, O., Duman, M., Güler, E. M."<p>Cytotoxic, apoptotic and genotoxic effects of thymoquinone‑oxime derivative on gastric cancer cells: An <em>in vitro</em> study</p>". Oncology Letters 31, no. 3 (2026): 109. https://doi.org/10.3892/ol.2026.15462
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