Background: Hepatocellular carcinoma stem cells (HCSCs) contribute to tumor recurrence and therapy resistance, necessitating novel therapeutic strategies. Artesunate (ART), an anti-malarial drug, has demonstrated anti-cancer effects, but its mechanism in HCSCs remains unclear. Methods: CD133 + Hep3B cells were treated with ART, and cell viability was assessed via CCK-8 assay. Ferroptosis-related pathways were analyzed using RNA sequencing and KEGG analysis. Key ferroptosis markers and oxidative stress indicators were tested. The ferroptosis inhibitor deferoxamine (DFO) was used to validate the mechanism. Stemness (spheroid formation), malignancy (Ki67, HSP90), and metastatic potential (migration, invasion) were examined in vitro . Tumorigenesis was assessed and ART-inducing ferroptosis mechanism was further validated in xenograft models. Results: ART dose- and time-dependently inhibited HCSCs proliferation (IC50: 82.35 μM at 12 h, 34.97 μM at 24 h and 19.76 μM at 48 h, respectively). RNA sequencing revealed ferroptosis as the primary pathway. ART upregulated TFRC, FTL and FTH1 while increasing ferric ions, ROS and MDA effects reversed by DFO. ART suppressed stemness, malignancy and metastatic potential in vitro . In vivo , ART reduced tumor growth, which was rescued by DFO, confirming iron metabolism-related ferroptosis dependence. Conclusion: ART induced ferroptosis in HCSCs by disrupting iron metabolism, leading to oxidative stress and lipid peroxidation. This mechanism underlies its suppression of stemness, proliferation, and metastasis, highlighting ART's potential as a targeted therapy for hepatocellular carcinoma (HCC).
Nie et al. (2026) studied this question.