Linghu Lingjuan, Zhou Hongying, Zheng Gang, Yi Tao
Development and Related Diseases of Women and Children Key Laboratory of Sichuan Province, Key Laboratory of Birth Defects and Related Diseases of Women and Children, Ministry of Education, West China Second Hospital, Sichuan University, Chengdu, 610041, China.
Department of Human Anatomy, West China School of Preclinical and Forensic Medicine, Sichuan University, Chengdu, 610041, China.
Oncol Res. 2025 Aug 28;33(9):2421-2434. doi: 10.32604/or.2025.063724. eCollection 2025.
Ovarian cancer, a leading cause of gynecological malignancy-related mortality, is characterized by limited therapeutic options and a poor prognosis. Although pyrimethamine has emerged as a promising candidate demonstrating efficacy in treating various tumors, the precise mechanisms of its antitumor effects remain obscure. This study was specifically designed to investigate the mode of action underlying the antitumor effects of pyrimethamine in preclinical settings.
The effects of pyrimethamine on cellular proliferation were meticulously assessed using both the cell counting kit 8 (CCK-8) assay and the colony formation assay, with the effects further confirmed in a murine model. A confocal microscope was utilized to monitor the dynamic alterations in mitochondria within ovarian cancer cells. Additionally, adenosine triphosphate (ATP) and reactive oxygen species (ROS) assays were conducted to measure mitochondrial damage induced by pyrimethamine in ovarian cancer cell lines. The mitochondrial membrane potential was assessed using fluorescent dyes as an indicator of mitochondrial functional status. Furthermore, transcriptome analysis and immunohistochemical techniques were employed to detect the impact of pyrimethamine on ovarian cancer cells.
Our results demonstrated that pyrimethamine induced ovarian cancer cell death through mitochondrial dysfunction and lethal mitophagy. Transcriptome profiling analysis and Western blot demonstrated that activation of the p38/JNK/ERK signaling pathway was implicated in the process of pyrimethamine-induced mitophagy in ovarian cancer cells. Importantly, combination treatment with pyrimethamine and paclitaxel and showed a synergistic antitumor effect.
Altogether, these findings indicate that the antitumor effects of pyrimethamine result from the induction of lethal mitophagy via regulation of the p38/JNK/ERK pathway in ovarian cancer. Considering the low toxicity and high tolerance associated with pyrimethamine, it is suggested that pyrimethamine be evaluated in the treatment of ovarian cancer, either as a monotherapy or in combination with paclitaxel.
卵巢癌是妇科恶性肿瘤相关死亡的主要原因，其特点是治疗选择有限且预后不良。尽管乙胺嘧啶已成为一种有前景的候选药物，在治疗各种肿瘤方面显示出疗效，但其抗肿瘤作用的确切机制仍不清楚。本研究专门设计用于在临床前环境中研究乙胺嘧啶抗肿瘤作用的作用模式。
使用细胞计数试剂盒8（CCK-8）测定法和集落形成测定法精心评估乙胺嘧啶对细胞增殖的影响，并在小鼠模型中进一步证实其效果。利用共聚焦显微镜监测卵巢癌细胞中线粒体的动态变化。此外，进行了三磷酸腺苷（ATP）和活性氧（ROS）测定，以测量乙胺嘧啶在卵巢癌细胞系中诱导的线粒体损伤。使用荧光染料评估线粒体膜电位，作为线粒体功能状态的指标。此外，采用转录组分析和免疫组织化学技术检测乙胺嘧啶对卵巢癌细胞的影响。
我们的结果表明，乙胺嘧啶通过线粒体功能障碍和致死性线粒体自噬诱导卵巢癌细胞死亡。转录组分析和蛋白质印迹表明，p38/JNK/ERK信号通路的激活与乙胺嘧啶诱导卵巢癌细胞线粒体自噬的过程有关。重要的是，乙胺嘧啶与紫杉醇联合治疗显示出协同抗肿瘤作用。
总之，这些发现表明，乙胺嘧啶的抗肿瘤作用是通过调节卵巢癌中的p38/JNK/ERK途径诱导致死性线粒体自噬而产生的。考虑到乙胺嘧啶的低毒性和高耐受性，建议对乙胺嘧啶进行评估，用于卵巢癌的治疗，可作为单一疗法或与紫杉醇联合使用。
