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Investigation of cellular effects of thymoquinone on glioma cell
Eray Metin Guler1,2,3, Behice Hande Sisman4, Abdurrahim Kocyigit2,3
1Department of Medical Biochemistry, Faculty of Hamidiye Medicine, University of Health Sciences Turkey, Istanbul, Turkey.
Abstract:
Glioblastoma, as an invasive tumor, is one of the most common primary malignant brain tumors. Despite maximum aggressive treatment, patients with glioblastoma have a dismal prognosis. Thymoquinone (TQ) has been found to show anti-cancer effects on different types of cancer. There are a few in vitro studies on the effect of TQ on glial tumors. However, the molecular mechanism of TQ's anti-cancer effect has not been fully elucidated. In the present study, we aimed to investigate the genotoxic, apoptotic, and cytotoxic effects of TQ on C6 rat glioma cells. C6 glioma cells were analyzed after 24 h of exposure to different concentrations of TQ by the ATP cell viability assay for cytotoxicity, comet assay for genotoxicity, 2',7'dichlorodihydrofluorescein diacetate (H2DCF-DA) for intracellular reactive oxygen species (iROS) generation, 3.3'dihexyloxacarbocyanine iodide (DiOC6(3)) for mitochondrial membrane potential, GSH/GSSG-Glo Assay for glutathione level and Fura-2AM for intracellular calcium levels. Apoptosis induction was studied by acridine orange/ethidium bromide double staining, flow cytometry, and western blotting analyses. Caspase-3, Caspase-9, Bax, Bcl-2, and pSTAT3 protein levels were determined by the western blotting method. Cytotoxicity was enhanced by TQ in C6 glioma cells in a concentration-dependent manner. TQ also induced DNA damage, apoptosis, and increased iROS. Also, MMP and GSH levels were decreased by TQ. It inhibited pSTAT3, resulting in apoptosis induction through the regulation of anti-apoptotic and pro-apoptotic proteins. Our results suggest that TQ would be an effective treatment in glioma. Further studies should support these findings.
Insights
Thymoquinone (TQ) exhibits anti-cancer properties against glioma cells by inducing DNA damage, apoptosis, and reactive oxygen species. This natural compound shows potential as an effective treatment for glioma, warranting further investigation.
Area of Science:
- * Oncology
- * Molecular Biology
- * Pharmacology
Background:
- * Glioblastoma is an aggressive brain tumor with a poor prognosis despite intensive treatments.
- * Thymoquinone (TQ) demonstrates anti-cancer effects, but its precise molecular mechanisms in glioma remain unclear.
- * Existing research on TQ's effects on glial tumors is limited, necessitating further investigation.
Purpose of the Study:
- * To investigate the genotoxic, apoptotic, and cytotoxic effects of Thymoquinone (TQ) on C6 rat glioma cells.
- * To elucidate the molecular mechanisms underlying TQ's anti-cancer activity in glioma.
- * To evaluate TQ's impact on intracellular reactive oxygen species (iROS), mitochondrial membrane potential (MMP), glutathione (GSH) levels, and calcium signaling.
Main Methods:
- * Cytotoxicity assessed via ATP cell viability assay.
- * Genotoxicity evaluated using the comet assay.
- * iROS, MMP, GSH, and calcium levels measured using specific assays (H2DCF-DA, DiOC6(3), GSH/GSSG-Glo, Fura-2AM).
- * Apoptosis induction analyzed through acridine orange/ethidium bromide staining, flow cytometry, and Western blotting.
- * Protein levels of Caspase-3, Caspase-9, Bax, Bcl-2, and pSTAT3 determined by Western blotting.
Main Results:
- * TQ demonstrated a dose-dependent increase in cytotoxicity against C6 glioma cells.
- * TQ induced significant DNA damage, apoptosis, and elevated intracellular reactive oxygen species (iROS).
- * TQ treatment led to decreased mitochondrial membrane potential (MMP) and glutathione (GSH) levels.
- * TQ inhibited pSTAT3 signaling, modulating pro-apoptotic (Bax) and anti-apoptotic (Bcl-2) proteins to promote apoptosis.
Conclusions:
- * Thymoquinone exhibits potent cytotoxic, genotoxic, and apoptotic effects on C6 rat glioma cells.
- * TQ's mechanism involves iROS generation, MMP disruption, and pSTAT3 inhibition, leading to apoptosis.
- * TQ shows promise as a potential therapeutic agent for glioma treatment, meriting further clinical research.
