Artesunate induces ferroptosis in osteosarcoma through <scp>NCOA4</scp> ‐mediated ferritinophagy

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 Fe²⁺ 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.