Exploration of the pharmacological mechanism of Evodia rutaecarpa on esophageal cancer based on comprehensive analysis of compounds and disease targets

Evodia rutaecarpa, a traditional Chinese medicine (TCM), possesses high medicinal value and has shown great potential in the treatment of tumors and Alzheimer's disease. However, its active components against esophageal cancer and the underlying molecular mechanisms remain unclear. In this study, Ultra Performance Liquid Chromatography-Orbitrap-Mass Spectrometry (UPLC-Orbitrap-MS) was used to analyze the chemical components of Evodia rutaecarpa. Key components were identified through TCMSP, SwissADME, and BATMAN-TCM databases, and target prediction was performed using SwissTargetPrediction. Potential therapeutic targets were retrieved from OMIM, GeneCards, and TTD databases. Cytoscape and STRING were used to construct a "medicine-component-target" network, followed by GO/KEGG enrichment analysis. Molecular docking and dynamics simulations were performed with Schrödinger Maestro. In vitro validation was done using CCK-8, EdU, IF assays, flow cytometry, and Western blot analysis on esophageal squamous cell carcinoma cells. Through this study, 13 active components and 127 potential targets were identified. GO/KEGG analysis revealed key roles in cell cycle regulation. 6-hydroxyluteolin, isorhamnetin, and rutaecarpine were identified as major active components. Molecular dynamics simulations confirmed strong binding to key targets (CDK2, SRC, EGFR). In vitro experiments demonstrated that 6-hydroxyluteolin and isorhamnetin significantly inhibited cell proliferation, upregulated γ-H2AX expression levels, induced cell cycle arrest, and ultimately induced apoptosis. We found that Evodia rutaecarpa exerts anti-esophageal cancer effects through multiple components, targets, and pathways, thereby providing a theoretical basis for the potential application of Evodia rutaecarpa in the treatment of esophageal cancer.