Integrated analysis of Morusin inhibiting esophageal cancer via network pharmacology molecular dynamics simulation and in vitro validation

Esophageal cancer (EC) is a malignant tumor characterized by high incidence and mortality, and its complex pathogenesis necessitates the development of effective therapeutic strategies. In this study, we investigated the anti-cancer effects and underlying mechanisms of Morusin against EC using network pharmacology, molecular dynamics (MD) simulation, and in vitro experiments. Transcriptomic analysis identified 3,540 differentially expressed genes in EC as potential therapeutic targets, while potential targets of Morusin were predicted using the SWISSprediction database. The intersection analysis revealed 21 common genes, which may represent the key targets of Morusin in EC treatment. A protein–protein interaction (PPI) network was constructed using the STRING database and analyzed in Cytoscape 3.8.0, identifying MAPK3, PTGS2, and HSP90AA1 as core hub genes. GO enrichment analysis indicated that the biological processes involved in Morusin’s anti-EC activity mainly included peptidyl-serine phosphorylation, peptidyl-serine modification, and aging. The cellular components were primarily associated with focal adhesion, cell–substrate junction, and caveola, while the molecular functions were enriched in protein serine kinase activity, protein serine/threonine kinase activity, protein serine/threonine/tyrosine kinase activity, and MAP kinase kinase activity. KEGG pathway enrichment suggested that Morusin may exert its effects through the HIF-1 signaling pathway, prostate cancer, and serotonergic synapse pathways. Molecular docking and MD simulations demonstrated stable binding of Morusin to ERK1 (MAPK3) and MEK1. In vitro assays revealed that 10 µM Morusin markedly inhibited the proliferation of EC cells (KYSE150 and TE1) without significant cytotoxicity toward normal esophageal epithelial cells (Het-1 A). Furthermore, Transwell and wound-healing assays confirmed that Morusin suppressed EC cell migration and invasion. Western blot analysis showed that Morusin treatment decreased Bcl-2 expression and increased cleaved-caspase3/caspase3, cleaved-PARP/PARP, and Bax levels, accompanied by significant downregulation of p-MEK1/2, p-MAPK3, and p-ERK2. Collectively, these findings suggest that Morusin inhibits proliferation, migration, and invasion, and induces apoptosis in EC cells by suppressing the MAPK signaling pathway.