Unravelling the Mechanism of Methylophiopogonanone A Against Esophageal Squamous Cell Carcinoma Based on Network Pharmacology and Molecular Docking

INTRODUCTION: Esophageal squamous cell carcinoma (ESCC) stands as one of the deadliest cancers globally. Given the urgent clinical need for more precise and comprehen-sive therapeutic strategies, the phytocompound methylophiopogonanone A (MO-A) demon-strates the potential as a candidate for ESCC treatment. This study aimed to verify the ther-apeutic effect of MO-A against ESCC and unveil its underlying mechanism. METHODS: Three compound-protein interaction databases were utilized to predict the molec-ular targets of MO-A. Subsequently, potential therapeutic targets of ESCC were identified based on the GEO database. KEGG pathway and GO function enrichment analyses were then performed by using these two sets of targets, respectively. Through the integrative anal-ysis of these two target sets, core targets of MO-A with therapeutic potential against ESCC were determined. Protein-protein interaction network analyses and molecular dockings were executed by using these targets. Two human-derived ESCC cell lines were enrolled for bio-logical validation, including cell viability, colony formation, and cell cycle assays. RESULTS: This study predicted 200 potential targets of MO-A and uncovered 138 key targets associated with the progression of ESCC. Enrichment analyses and PPI networks under-scored the involvement of cell cycle-related genes in ESCC development. Four proteins were determined as core MO-A targets for ESCC treatment, including AURKA, AURKB, CDC25B, and TOP2A, which partake in the regulation of the cell cycle. Finally, the inhibi-tory effect of MO-A on ESCC cell proliferation was validated in vitro, primarily through inducing cell cycle arrest at the G2/M phase in ESCC cells. DISCUSSION: These results revealed the anti-ESCC potential of MO-A, a plant-derived fla-vonoid, using integrated bioinformatics and biological experiments. While findings provide a mechanistic basis for the efficacy of MO-A, limitations include reliance on computational and in vitro models. Further studies should be conducted to evaluate the pharmacological properties and safety of MO-A across multiple models, alongside more comprehensive structure-activity relationship studies to inform drug optimization prior to clinical transla-tion. CONCLUSION: MO-A can impede ESCC growth by triggering cell cycle G2/M arrest, posi-tioning it as a novel and promising phytocompound for ESCC therapy.