Huang Rui, Xu Ruiqing, Shi Jiandang, Yang Zongqiang, Zheng Jianping, Wei Daihao
Department of Orthopedic Surgery, General Hospital of Ningxia Medical University, Yinchuan, China.
The First School of Clinical Medicine, General Hospital of Ningxia Medical University, Yinchuan, China.
FASEB J. 2025 Apr 15;39(7):e70488. doi: 10.1096/fj.202403160R.
Osteosarcoma (OS) is a prevalent primary malignant bone tumor that lacks effective therapeutic interventions. Artesunate (ART) has been proved to have remarkable treatment effects on severe malaria and anti-tumor properties. This study aimed to investigate the anti-OS effects and underlying mechanisms of ART. The potential mechanisms of ART-mediated anti-OS activity were analyzed by using RNA sequencing, iron accumulation, lipid peroxidation, western blotting, and small interfering RNA (siRNA) transfection. In vivo, a xenograft mice model was adopted to explore the anticancer effect of ART. The present study revealed that ART significantly suppressed OS cell proliferation. Subsequent results suggested that ART exerted anti-OS activity mainly through the ferroptosis pathway. ART decreased the GSH/GSSG ratio, xCT and GPX4 expression, while increasing MDA and lipid peroxidation, which were reversed by Fer-1, DFO, 3-MA, and NCOA4 silencing. Mechanistically, ART upregulated the expression of TFR and DMT1, and triggered ferritinophagy by upregulating the expression of NCOA4, which increased Fe accumulation and triggered ferroptosis. In addition, cytoplasmic iron further activated Mfrn2-mediated transportation of cytoplasmic free iron into the mitochondria, resulting in mitochondrial iron overload, eventually leading to lipid peroxidation and ferroptosis. Furthermore, in an OS xenograft mouse model, administration of ART inhibited tumor growth by ferroptosis. Collectively, our findings indicated that ART has the potential anti-OS capacity through NCOA4-mediated ferritinophagy, which might shed light on the future of OS therapy.
骨肉瘤（OS）是一种常见的原发性恶性骨肿瘤，缺乏有效的治疗干预措施。青蒿琥酯（ART）已被证明对重症疟疾具有显著的治疗效果和抗肿瘤特性。本研究旨在探讨ART的抗OS作用及其潜在机制。通过RNA测序、铁蓄积、脂质过氧化、蛋白质免疫印迹和小干扰RNA（siRNA）转染分析了ART介导的抗OS活性的潜在机制。在体内，采用异种移植小鼠模型来探究ART的抗癌作用。本研究表明，ART显著抑制OS细胞增殖。随后的结果表明，ART主要通过铁死亡途径发挥抗OS活性。ART降低了谷胱甘肽/氧化型谷胱甘肽比值、xCT和谷胱甘肽过氧化物酶4（GPX4）的表达，同时增加了丙二醛（MDA）和脂质过氧化，而铁抑素-1（Fer-1）、去铁胺（DFO）、3-甲基腺嘌呤（3-MA）和核受体辅激活因子4（NCOA4）沉默可逆转这些变化。机制上，ART上调了转铁蛋白受体（TFR）和二价金属离子转运体1（DMT1）的表达，并通过上调NCOA4的表达引发铁自噬，从而增加铁蓄积并引发铁死亡。此外，细胞质铁进一步激活线粒体铁转运蛋白2（Mfrn2）介导的细胞质游离铁向线粒体的转运，导致线粒体铁过载，最终导致脂质过氧化和铁死亡。此外，在OS异种移植小鼠模型中，给予ART可通过铁死亡抑制肿瘤生长。总的来说，我们的研究结果表明，ART具有通过NCOA4介导的铁自噬发挥潜在抗OS的能力，这可能为OS治疗的未来发展提供思路。
