Middle East Journal of Cancer

Exploring the Apoptotic and Cytotoxic Effects of Thymol in C6 Glioma Cells

Document Type : Original Article(s)

Authors

Negar Ghazvini 1
Pejman Hosseinzadeh 2
Sadaf Afshari 2
Bahram Bibak 3
Mahsa Jalali 4
Ghazale Biglari 2
Seyed Sajad Ahmadi 5
Reza Salarinia 3
Amir R. Afshari 3, 6, 7

1 Student Research Committee, North Khorasan University of Medical Sciences, Bojnurd, Iran

2 Student Research Committee, Mashhad University of Medical Sciences, Mashhad, Iran

3 Natural Products and Medicinal Plants Research Center, North Khorasan University of Medical Sciences, Bojnurd, Iran

4 Faculty of Medicine, Mashhad University of Medical Sciences, Mashhad, Iran

5 Department of Ophthalmology, Khatam-Ol-Anbia Hospital, Mashhad University of Medical Sciences, Mashhad, Iran

6 Department of Basic Sciences, Faculty of Medicine, Mashhad Medical Sciences, Islamic Azad University, Mashhad, Iran

7 Department of Physiology and Pharmacology, Faculty of Medicine, North Khorasan University of Medical Sciences, Bojnurd, Iran

https://doi.org/10.30476/mejc.2026.106361.2261
Abstract
Background: Glioblastoma (GBM) is the most common and aggressive brain tumor associated with a poor prognosis, highlighting the need for new therapeutic strategies. The present study aimed to evaluate the anticancer properties of thymol in rat GBM C6 malignant cells.
Method: This in-vitro study was conducted to evaluate the effects of thymol on cell viability and apoptosis using the resazurin assay and Annexin V-FITC/PI staining kit, respectively. Moreover, reactive oxygen species (ROS) generation was evaluated using the Dichlorodihydrofluorescein diacetate assay, and the mRNA levels of Bax, Bcl2, and p53 were assessed using quantitative real-time polymerase chain reaction. Data from three independent experiments were analyzed using GraphPad Prism 8.2.1 and compared for statistically significant differences (P < 0.05) using a one-way ANOVA with a Tukey post-hoc test.
Results: The half-maximal inhibitory concentration (IC50) of thymol for C6 cells was 350 μM at 24 h and 260 μM at 48 h post-treatment. At the same time, lower cytotoxic effects were observed on normal human foreskin fibroblast cells (IC50 = 675 μM). 350 μM of thymol also caused a significant G2/M cell cycle arrest and early and late apoptosis compared with the non-treated control group. Thymol also caused a significant increase in Bax and p53 expression, as well as ROS generation, when cells were treated with thymol compared with the non-treated control group.
Conclusion: The results indicate the anticancer properties of thymol including cytotoxic, and pro-oxidant effects with selectivity towards cancer cells. These preclinical findings provide a strong rationale for further exploration of thymol as a potential phytochemical agent in combating GBM.

Highlights

Amir R. Afshari (google scholar)

Reza Salarinia (google scholar)

Keywords

Glioblastoma multiforme
Thymol
Cancer
C6 cell line
Apoptosis
Subjects

Please cite this article as:

Ghazvini N, Hosseinzadeh P, Afshari S, Bibak B, Jalali M, Biglari GH, et al. Exploring the apoptotic and cytotoxic effects of thymol in C6 glioma cells. Middle East J Cancer. 2026; 17(4): 285-96. doi: 10.30476/mejc.2026. 106361.2261.

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