Guler Eray Metin, Sisman Behice Hande, Kocyigit Abdurrahim, Hatiboglu Mustafa Aziz
Department of Medical Biochemistry, Faculty of Hamidiye Medicine, University of Health Sciences Turkey, Istanbul, Turkey.
Department of Medical Biochemistry, Bezmialem Vakif University, Faculty of Medicine, Istanbul, Turkey.
Toxicol Rep. 2021 Jan 4;8:162-170. doi: 10.1016/j.toxrep.2020.12.026. eCollection 2021.
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 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 (HDCF-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.
胶质母细胞瘤作为一种侵袭性肿瘤，是最常见的原发性恶性脑肿瘤之一。尽管进行了最大程度的积极治疗，胶质母细胞瘤患者的预后仍然很差。已发现百里醌（TQ）对不同类型的癌症具有抗癌作用。关于TQ对胶质瘤影响的研究较少。然而，TQ抗癌作用的分子机制尚未完全阐明。在本研究中，我们旨在研究TQ对C6大鼠胶质瘤细胞的遗传毒性、凋亡和细胞毒性作用。通过ATP细胞活力测定法检测细胞毒性、彗星试验检测遗传毒性、2',7'-二氯二氢荧光素二乙酸酯（HDCF-DA）检测细胞内活性氧（iROS）生成、3,3'-二己基氧杂羰花青碘化物（DiOC6(3)）检测线粒体膜电位、GSH/GSSG-Glo检测法检测谷胱甘肽水平以及Fura-2AM检测细胞内钙水平，对暴露于不同浓度TQ 24小时后的C6胶质瘤细胞进行分析。通过吖啶橙/溴化乙锭双重染色、流式细胞术和蛋白质印迹分析研究凋亡诱导情况。通过蛋白质印迹法测定半胱天冬酶-3、半胱天冬酶-9、Bax、Bcl-2和pSTAT3蛋白水平。TQ以浓度依赖性方式增强了C6胶质瘤细胞的细胞毒性。TQ还诱导了DNA损伤、凋亡并增加了iROS。此外，TQ降低了MMP和GSH水平。它抑制pSTAT3，通过调节抗凋亡和促凋亡蛋白诱导凋亡。我们的结果表明，TQ可能是治疗胶质瘤的有效方法。进一步的研究应支持这些发现。
