The Mechanism of Ranunculus ternatus Thunb. against Esophageal Squamous Cell Carcinoma Based on Network Pharmacology and Experimental Verification.

Background: Esophageal Squamous Cell Carcinoma (ESCC) remains a significant global health challenge, underscoring the urgent need for the development of innovative therapeutic approaches. Ranunculus ternatus Thunb., a traditional herbal medicine, has shown potential anticancer properties, but its pharmacological mechanisms against ESCC remain poorly understood. This study aimed to elucidate the molecular mechanisms of the ethyl acetate extract of Ranunculus ternatus Thunb. (RTE) against ESCC by integrating network pharmacology analysis with experimental validation.

Methods: Potential targets of RTE and ESCC were identified through network pharmacology using public databases. A Protein-Protein Interaction (PPI) network was constructed to identify key targets, followed by Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway enrichment analyses to explore biological processes and signaling pathways. The predicted mechanisms were experimentally validated using in vitro assays, including cell proliferation analysis and western blot assay in ESCC cell lines.

Results: Network pharmacology analysis identified 274 potential targets, with 14 key genes implicated in the therapeutic effects of RTE. GO analysis revealed significant involvement in the inflammatory response and apoptotic signaling pathways. KEGG pathway analysis highlighted the MAPK, Relaxin, and PI3K/Akt signaling pathways as critical mechanisms. In vitro experiments demonstrated that RTE significantly inhibited the proliferation of EC-109 and TE-13 cells by modulating the MAPK/ERK and PI3K/Akt pathways.

Conclusion: This study provides a comprehensive understanding of the molecular mechanisms underlying the anticancer effects of RTE against ESCC. These findings underscore the potential of RTE as a promising natural compound for ESCC treatment.