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Anticancer effects of thymoquinone on prostate cancer cells and natural killer cell activity

  • Authors:
    • Nouf A. Aldarmahi
    • Nesrin I. Tarbiah
    • Nuha A. Alkhattabi
    • Huda F. Alshaibi
  • View Affiliations / Copyright

    Affiliations: Department of Biochemistry, Faculty of Science, King Abdulaziz University, Jeddah 21589, Saudi Arabia
    Copyright: © Aldarmahi et al. This is an open access article distributed under the terms of Creative Commons Attribution License.
  • Article Number: 530
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    Published online on: September 18, 2025
       https://doi.org/10.3892/ol.2025.15276
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Abstract

Dark cumin, or Nigella sativa, is a well‑known and popular herbal medicine that may be used in the treatment of gastrointestinal issues, diabetes, bronchitis, high blood pressure and cancer. The anticancer action of Nigella sativa may be associated with the immune‑modulatory effect of its activation of human natural killer (NK) cells. NK cells, a subset of lymphocytes, are a component of innate immunity, the body's initial line of defense against infections. To identify potential ways to improve immunotherapy using natural products, the present study aimed to assess the impact of thymoquinone (TQ), a key component of Nigella sativa, on the activity of NK cells through their cytotoxicity and released chemicals. CD56 NK cells that were isolated and positively selected from blood samples, were cultured with PC3‑RFP prostate cancer cells and treated with TQ at different concentrations (25 and 50 µM). Cytotoxicity was measured, and perforin, granzyme B and interferon‑α (IFN‑α) levels were assessed using ELISA. The results demonstrated that the cytotoxic effect of NK cells on PC3‑RFP was significantly increased in the presence of TQ from 55.1 to 85.35%. Furthermore, with 50 µM TQ, there was a significant increase in secretion of perforin (0.80 ng/ml) compared with the control (0.2 ng/ml), and of granzyme B (1.24 ng/ml) compared with the control (0.65 ng/ml). Moreover, the production of IFN‑α by the NK cells significantly increased in the presence of 25 and 50 µM TQ (0.5 and 0.7 ng/ml, respectively) compared with the control (0.3 ng/ml). In summary, TQ increased NK cell cytotoxicity against PC3‑RFP cells, indicating a potential application in the development of cell‑based immunity treatment that requires more research and has promising prospects. 
View Figures

Figure 1

Cytotoxicity mechanism of NK cells.
By evaluating the net input of activating and inhibitory signals
from target cells through NK cell surface receptors, NK cells
discriminate healthy cells from target cells. However, several
tumors develop mechanisms to escape NK cell immune responses by
modifying their cell surface molecules, such as low immunogenicity.
Following the identification of the target cell by NK cells, the
stimulation domain is activated. This is initiated by the secretion
of a cytolytic enzyme, known as perforin, stored in NK cell
vesicles. Perforin creates pores in the cell membrane of the target
cell, allowing the passage of granzymes into the target cell and
starting protein digestion (cytolysis). NK cells express TRAIL and
FasL, which interact with TRAIL receptors and FAS ligands in cancer
cells, respectively. This interaction results in cell death,
signaling complex stimulating apoptosis. NK, natural killer; TRAIL,
TNF-related apoptosis inducing ligand; TRAILR, TRAIL receptor; MHC,
major histocompatibility complex; FasL, FAS ligand. Created in
BioRender.com.

Figure 2

NK cells dot plot before treatment.
CD56 positivity was determined by comparison with defined cut-off
values obtained from unstained control cells. The NK cell purity
was 95.7%. NK, natural killer.

Figure 3

NK cell cytotoxicity against PC3-RFP
cells in the presence or absence of TQ. The effector (NK cells) and
target (PC3-RFP) ratio was 1:2. *P<0.05; **P<0.01;
***P<0.001. NK, natural killer; TQ, thymoquinone; TC, tumor
cells.

Figure 4

Production of major cytokines against
PC3-RFP cells by NK cells in the presence or absence of TQ. TQ
enhances the release of (A) perforin, (B) granzyme B and (C) IFN-α,
the major secreted cytokines of NK cells. *P<0.05; **P<0.01;
***P<0.001. NK, natural killer; TQ, thymoquinone; TC, tumor
cells.

Figure 5

Spearman correlation coefficients
(2-tailed) of perforin, IFN-α and granzyme B cytokines, and natural
killer cytotoxicity with PC3-RFP cells.
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Copy and paste a formatted citation
Spandidos Publications style
Aldarmahi NA, Tarbiah NI, Alkhattabi NA and Alshaibi HF: Anticancer effects of thymoquinone on prostate cancer cells and natural killer cell activity. Oncol Lett 30: 530, 2025.
APA
Aldarmahi, N.A., Tarbiah, N.I., Alkhattabi, N.A., & Alshaibi, H.F. (2025). Anticancer effects of thymoquinone on prostate cancer cells and natural killer cell activity. Oncology Letters, 30, 530. https://doi.org/10.3892/ol.2025.15276
MLA
Aldarmahi, N. A., Tarbiah, N. I., Alkhattabi, N. A., Alshaibi, H. F."Anticancer effects of thymoquinone on prostate cancer cells and natural killer cell activity". Oncology Letters 30.5 (2025): 530.
Chicago
Aldarmahi, N. A., Tarbiah, N. I., Alkhattabi, N. A., Alshaibi, H. F."Anticancer effects of thymoquinone on prostate cancer cells and natural killer cell activity". Oncology Letters 30, no. 5 (2025): 530. https://doi.org/10.3892/ol.2025.15276
Copy and paste a formatted citation
x
Spandidos Publications style
Aldarmahi NA, Tarbiah NI, Alkhattabi NA and Alshaibi HF: Anticancer effects of thymoquinone on prostate cancer cells and natural killer cell activity. Oncol Lett 30: 530, 2025.
APA
Aldarmahi, N.A., Tarbiah, N.I., Alkhattabi, N.A., & Alshaibi, H.F. (2025). Anticancer effects of thymoquinone on prostate cancer cells and natural killer cell activity. Oncology Letters, 30, 530. https://doi.org/10.3892/ol.2025.15276
MLA
Aldarmahi, N. A., Tarbiah, N. I., Alkhattabi, N. A., Alshaibi, H. F."Anticancer effects of thymoquinone on prostate cancer cells and natural killer cell activity". Oncology Letters 30.5 (2025): 530.
Chicago
Aldarmahi, N. A., Tarbiah, N. I., Alkhattabi, N. A., Alshaibi, H. F."Anticancer effects of thymoquinone on prostate cancer cells and natural killer cell activity". Oncology Letters 30, no. 5 (2025): 530. https://doi.org/10.3892/ol.2025.15276
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