Su Yu-Feng, Tsai Tai-Hsin, Kuo Keng-Liang, Wu Chieh-Hsin, Su Hui-Yuan, Chang Wen-Chang, Huang Fu-Long, Lieu Ann-Shung, Kwan Aij-Lie, Loh Joon-Khim, Lin Chih-Lung, Tsai Cheng Yu
Division of Neurosurgery, Department of Surgery, Kaohsiung Medical University Hospital, Kaohsiung, Taiwan.
Department of Surgery, Post Baccalaureate Medicine, College of Medicine, Kaohsiung Medical University, Kaohsiung, Taiwan.
PLoS One. 2025 May 9;20(5):e0320849. doi: 10.1371/journal.pone.0320849. eCollection 2025.
Toona sinensis, a kind of phytochemicals in traditional Chinese medicine widely used in South-East Asia, has been recognized for its anticancer properties, particularly against various types of cancer. We aimed to evaluate the effectiveness of T. sinensis leaf extracts (TSL) specifically for glioblastoma multiforme (GBM). Gallic acid was identified as the major active component in the aqueous extracts of TSL using the HPLC system. Furthermore, it has been shown to have the ability to penetrate the blood-brain barrier. Various concentrations of TSL (10, 20, 40, and 80 μg/mL) were applied and 80 μg/mL TSL treatment significantly inhibited cell growth, proliferation, and cytotoxicity in A172 and U251 GBM cells. Flow cytometry analysis revealed cell cycle arrest at the G2/M phase and increased apoptotic cells. Furthermore, we observed mitochondrial dysfunction characterized by elevated ROS levels and reduced ATP production due to the blockade of electron transport chain (ETC) complexes. TSL treatment regulated this ROS-induced mitochondrial dysfunction. Western blotting analysis showed upregulation of Bax and Puma, along with downregulation of Bcl-2. Additionally, TSL treatment induced the cleavage of caspase-3, caspase-9, and PARP, indicating activation of the mitochondria-mediated apoptosis pathway and caspase-dependent pathway in both GBM cell lines. To investigate the involvement of the MEK/ERK pathway in TSL-induced effects, we used U0126, an inhibitor of MEK/ERK kinase. The results demonstrated that TSL treatment suppressed MEK/ERK activation, inhibiting ROS-induced mitochondrial dysfunction and promoting apoptosis. This suggests a potential therapeutic strategy targeting the MEK/ERK pathway in GBM treatment. Overall, our findings indicate that TSL treatment exerts cytotoxic effects through ROS-mediated mitochondrial dysfunction and activation of apoptotic pathways via MEK/ERK pathway in GBM cells. These insights provide valuable knowledge for potential therapeutic applications of TSL in GBM treatment.
香椿是一种在东南亚广泛使用的中药植物化学物质，其抗癌特性，尤其是对各种类型癌症的抗癌特性已得到认可。我们旨在评估香椿叶提取物（TSL）对多形性胶质母细胞瘤（GBM）的有效性。使用高效液相色谱系统确定没食子酸是TSL水提取物中的主要活性成分。此外，已证明它具有穿透血脑屏障的能力。应用了不同浓度的TSL（10、20、40和80μg/mL），80μg/mL TSL处理显著抑制了A172和U251 GBM细胞的细胞生长、增殖和细胞毒性。流式细胞术分析显示细胞周期停滞在G2/M期，凋亡细胞增加。此外，我们观察到线粒体功能障碍，其特征是由于电子传递链（ETC）复合物的阻断导致活性氧水平升高和ATP生成减少。TSL处理调节了这种活性氧诱导的线粒体功能障碍。蛋白质印迹分析显示Bax和Puma上调，同时Bcl-2下调。此外，TSL处理诱导了caspase-3、caspase-9和PARP的裂解，表明在两种GBM细胞系中线粒体介导的凋亡途径和caspase依赖性途径均被激活。为了研究MEK/ERK途径在TSL诱导的效应中的作用，我们使用了MEK/ERK激酶抑制剂U0126。结果表明，TSL处理抑制了MEK/ERK激活，抑制了活性氧诱导的线粒体功能障碍并促进了凋亡。这表明在GBM治疗中靶向MEK/ERK途径的潜在治疗策略。总体而言，我们的研究结果表明，TSL处理通过活性氧介导的线粒体功能障碍和通过MEK/ERK途径激活GBM细胞中的凋亡途径发挥细胞毒性作用。这些见解为TSL在GBM治疗中的潜在治疗应用提供了有价值的知识。
