Vitexin attenuates gastric precancerous lesions with EGFR as a potential target based on network pharmacology and molecular simulations.

Precancerous gastric lesions (PLGC) represent a critical transitional stage toward gastric cancer, yet effective interventions to halt progression remain limited. Vitexin, a naturally occurring flavonoid, exhibits anti-cancer potential; however, its molecular targets and mechanisms in PLGC have not been fully defined. This study aimed to elucidate the targets and mechanisms of vitexin against PLGC using an integrative strategy combining network pharmacology, molecular docking/molecular dynamics (MD), and cellular validation. Putative targets of vitexin and PLGC-associated genes were intersected to obtain 33 common targets, followed by protein-protein interaction network construction and pathway enrichment analyses. Molecular docking was performed against core targets, and MD simulations were conducted to assess the stability of the EGFR-vitexin complex. In vitro validation was carried out in MNNG-induced MC cells using cell viability, migration, and invasion assays, phase-contrast imaging, and immunofluorescence staining of EGFR and E-cadherin. Network analysis highlighted CCND1, EGFR, and ABL1 as central nodes and implicated oncogenic programs including cell-cycle regulation and PI3K-Akt signaling pathway. Docking suggested favorable binding of vitexin to the core targets. MD simulations further supported a stable binding mode of vitexin within the EGFR pocket over 100 ns with preserved global compactness; a late-stage conformational adaptation accompanied by hydrogen-bond network reorganization was observed without signs of global unfolding. Functionally, vitexin reduced MC cell viability in a dose-dependent manner, reaching ~ 50% inhibition at 20 μM, and significantly suppressed cell migration and invasion. Immunofluorescence demonstrated decreased EGFR expression and altered membrane localization, together with restored E-cadherin expression and membrane localization, suggesting a shift toward an epithelial phenotype. Vitexin inhibits PLGC-associated malignant phenotypes through multi-target modulation. Among the identified candidates, EGFR may serve as a putative and functionally relevant target, while modulation of EGFR-associated signaling together with restoration of E-cadherin may contribute to the reversal of EMT-related phenotypes. These findings provide mechanistic and structural support for the further preclinical evaluation of vitexin in preventing gastric cancer progression.