# 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

## TL;DR

Broadleaf Vetch shows antitumor effects against esophageal cancer by blocking tumor spread and inducing cell death, suggesting it could be a new treatment option.

## Contribution

This study reveals Broadleaf Vetch's dual mechanism against esophageal cancer through EMT suppression and ferroptosis induction.

## Key findings

- Broadleaf Vetch inhibits epithelial–mesenchymal transition in esophageal cancer cells.
- The herb induces ferroptosis by increasing oxidative stress and depleting glutathione.
- Animal studies show reduced tumor growth and regulation of key cancer pathways.

## Abstract

Esophageal squamous cell carcinoma is an aggressive cancer with limited treatment options. In this study, we investigated Broadleaf Vetch (Vicia amoena), a traditional medicinal herb, and found that it shows antitumor effects against esophageal squamous cell carcinoma. By integrating computational network analysis with experimental verification, this study demonstrates that Broadleaf Vetch suppresses the epithelial–mesenchymal transition—a critical driver of tumor metastasis—while concurrently triggering ferroptosis, an iron-dependent form of regulated cell death characterized by oxidative stress. In animal studies, Broadleaf Vetch markedly reduced tumor growth and regulated key pathways associated with epithelial–mesenchymal transition and ferroptosis. These findings suggest Broadleaf Vetch as a potential natural multi-target candidate for the treatment of esophageal squamous cell carcinoma.

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.