Feng Qiao, Li Yuxi, Zhong Lincan, Nie Huwei, Yang Wensheng, Chen Guangyu, Zhang Lin
Department of Gastrointestinal Surgery, the Affiliated Hospital of Southwest Medical University, Luzhou, China.
Department of Gastrointestinal Surgery, General Hospital of Western Theater Command, Chengdu, China.
J Gastrointest Oncol. 2025 Jun 30;16(3):853-864. doi: 10.21037/jgo-2024-1002. Epub 2025 Jun 26.
Ferroptosis, a regulated form of cell death marked by iron-dependent lipid peroxidation, has gained recognition as a potential therapeutic target in diverse cancers, including gastrointestinal stromal tumors (GIST). The NCOA4-mediated ferritinophagy pathway is integral to the regulation of cellular iron homeostasis and the process of ferroptosis. Nevertheless, the effects of modulating this pathway on the viability of GIST cells and the induction of ferroptosis are yet to be elucidated. This study sought to examine the impact of toosendanin (TSN), a natural compound with prospective anticancer attributes, on ferroptosis in GIST cells, specifically emphasizing its regulatory influence on the NCOA4-mediated ferritinophagy pathway.
GIST-T1 cells were exposed to different concentrations of TSN. Cell viability, apoptosis, and ferroptosis were evaluated through flow cytometry (Annexin V/7-AAD), transmission electron microscopy (TEM), and biochemical detection. The proliferation, migration, and invasion capacities were assessed utilizing the Cell Counting Kit-8 (CCK-8) assay, clone formation assay, and Transwell assay, respectively. The impact of NCOA4 silencing and ferrostatin-1, an inhibitor of ferroptosis, was analyzed in conjunction with TSN treatment.
The results showed that TSN treatment markedly decreased cell viability and induced ferroptosis in GIST-T1 cells, as demonstrated by elevated levels of lipid reactive oxygen species (ROS), increased ferrous ion content, and membrane damage. Mechanistically, western blot analysis demonstrated that TSN downregulated the expression of key ferroptosis inhibitors glutathione peroxidase 4 (GPX4) and SLC7A11, while simultaneously upregulating NCOA4 and LC3II/I. The application of small interfering RNA (siRNA)-NCOA4 and ferrostatin-1, which inhibit NCOA4-mediated autophagy and ferroptosis, resulted in a significant restoration of GPX4 and SLC7A11 expression, thereby mitigating TSN-induced ferroptosis. Furthermore, TSN was found to effectively suppress the proliferation, migration, and invasion of GIST cells; these effects were reversed upon NCOA4 silencing and inhibition of ferroptosis.
This study underscores the potential of TSN as a therapeutic agent for GIST, particularly through the exploitation of NCOA4-mediated ferritinophagy to induce ferroptosis.
铁死亡是一种由铁依赖性脂质过氧化作用所标记的程序性细胞死亡形式，已被公认为包括胃肠道间质瘤（GIST）在内的多种癌症的潜在治疗靶点。NCOA4介导的铁蛋白自噬途径对于细胞铁稳态的调节和铁死亡过程至关重要。然而，调节该途径对GIST细胞活力和铁死亡诱导的影响尚待阐明。本研究旨在探讨具有潜在抗癌特性的天然化合物川楝素（TSN）对GIST细胞铁死亡的影响，特别强调其对NCOA4介导的铁蛋白自噬途径的调节作用。
将GIST-T1细胞暴露于不同浓度的TSN中。通过流式细胞术（Annexin V/7-AAD）、透射电子显微镜（TEM）和生化检测来评估细胞活力、凋亡和铁死亡。分别使用细胞计数试剂盒-8（CCK-8）检测、克隆形成检测和Transwell检测来评估细胞的增殖、迁移和侵袭能力。结合TSN处理分析NCOA4沉默和铁死亡抑制剂铁抑素-1的影响。
结果表明，TSN处理显著降低了GIST-T1细胞的活力并诱导了铁死亡，脂质活性氧（ROS）水平升高、亚铁离子含量增加和膜损伤证明了这一点。机制上，蛋白质印迹分析表明，TSN下调了关键铁死亡抑制剂谷胱甘肽过氧化物酶4（GPX4）和溶质载体家族7成员11（SLC7A11）的表达，同时上调了NCOA4和微管相关蛋白1轻链3II/I（LC3II/I）。应用抑制NCOA4介导的自噬和铁死亡的小干扰RNA（siRNA）-NCOA4和铁抑素-1导致GPX4和SLC7A11表达显著恢复，从而减轻了TSN诱导的铁死亡。此外，发现TSN能有效抑制GIST细胞的增殖、迁移和侵袭；在NCOA4沉默和铁死亡抑制后，这些作用被逆转。
本研究强调了TSN作为GIST治疗药物的潜力，特别是通过利用NCOA4介导的铁蛋白自噬来诱导铁死亡。
