Mechanism and molecular targets of Euphorbia fischeriana Steud root extract in hepatocellular carcinoma

Ethnopharmacological relevance Euphorbia fischeriana Steud. (E. fischeriana) root is a well-known traditional Chinese medicine used in the treatment of skin ulceration, lymph node tuberculosis and tumors. However, its antitumor activity against HCC and the underlying molecular mechanisms remain to be further elucidated. Aim of the study The study aimed to investigate the anti-hepatocarcinogenic effects of E. fischeriana root, specifically its impact on NCOA4-mediated ferritinophagy, and to elucidate the related molecular mechanisms in HCC. Methods LC-MS was used to analyze the comprehensive chemical composition of E. fischeriana root extract. Through network pharmacology, transcriptomics, molecular docking, and molecular dynamics simulations, we investigated the potential mechanisms underlying the effects of E. fischeriana root extract on HCC. Finally, in vitro and in vivo experiments were performed to validate these mechanisms. Results Through mass spectrometry analysis and drug-likeness screening, 93 active compounds were identified. Based on network pharmacology and transcriptomic analyses, we hypothesized that the PI3K/AKT pathway and its downstream signaling involving autophagy and ferroptosis are crucial pathways affected by E. fischeriana root extract. In vitro experiments demonstrated that E. fischeriana root extract inhibits proliferation, promotes apoptosis, triggers ferritinophagy and induces ferroptosis in HCC cells. In vivo studies further validated significant anti-HCC effects of E. fischeriana root extract. Conclusions Based on these findings, we propose that E. fischeriana root extract exerts its anti-HCC effects by targeting the PI3K/AKT pathway to regulate NCOA4-mediated ferritinophagy, thereby inducing ferroptosis. These results highlight E. fischeriana root extract as a promising therapeutic candidate for HCC.