Antitumor Effects of Broadleaf Vetch Against Esophageal Squamous Cell Carcinoma Through Dual Mechanisms: Suppressing EMT and Inducing Ferroptosis with Predicted Hepatorenal Toxicity-An Integrative Network Pharmacology and Toxicology Study

Background: Esophageal squamous cell carcinoma (ESCC) remains a highly lethal malignancy with limited effective treatments. Broadleaf Vetch (Vicia amoena, BV) is a traditional medicinal herb with potential Anticancer properties, but its mechanisms in ESCC are not fully understood.

Methods: Network pharmacology was used to identify BV-related therapeutic targets and pathways. Molecular docking validated interactions between BV components and core proteins. In vitro assays evaluated proliferation, colony formation, migration, invasion, epithelial-mesenchymal transition (EMT) markers, and ferroptosis-related indices. An ESCC xenograft model was used to assess antitumor efficacy in vivo.

Results: Five major BV components and 363 ESCC-related targets were identified, highlighting the PI3K-AKT pathway and key nodes such as EGFR, Akt1, Src, TP53, and GPX4. BV significantly inhibited ESCC cell proliferation, migration, and invasion, and reversed EMT marker expression. Ferroptosis induction was evidenced by significant Fe²⁺ accumulation, elevated Reactive Oxygen Species (ROS) and malondialdehyde levels, alongside glutathione depletion. BV treatment also precipitated mitochondrial dysfunction. In parallel, BV downregulated GPX4 and SLC7A11. Notably, these changes were largely reversed by the Ferroptosis inhibitor Ferrostatin-1. In vivo, BV suppressed tumor growth and regulated EMT- and ferroptosis-associated proteins in xenograft tissues.

Conclusions: BV exerts dual antitumor effects in ESCC by suppressing EMT and inducing Ferroptosis. These findings suggest BV may represent a potential multi-target phytotherapeutic candidate for ESCC.

broadleaf vetch; esophageal squamous cell carcinoma; ferroptosis; hepatotoxicity; nephrotoxicity; network pharmacology.