Objective: Anthriscus sylvestris (AS), a traditional medicinal herb, has emerged as a potential anti-cancer functional food. This study aimed to explore its anti-tumor mechanisms, focusing on colon cancer, using network pharmacology and experimental validation.

Materials and Methods: Active components of Anthriscus sylvestris were screened via the TCMSP database (OB≥30%). Network pharmacology identified 91 shared targets between Anthriscus sylvestris and anti-tumor activity. Protein-protein interaction (PPI) networks, GO/KEGG analyses, and molecular docking were performed. In vitro experiments assessed the effects of Anthriscus sylvestris petroleum ether extract on STAT3 activity in HT29 colon cancer cells.

Results: Three key active components (dihydroanhydropodorhizol, dimethylmatairesinol, and (+)-hinokinin) were identified. Molecular docking revealed strong binding affinities (e.g., −7.84 kcal/mol for (+)-hinokinin-ESR1). KEGG enrichment highlighted pathways like FoxO and STAT3 signaling. In HT29 cells, AS extract concentration and time-dependently suppressed STAT3/p-STAT3 protein expression (e.g., 80 μg/ml reduced STAT3 to 21.37% of control) and inhibited STAT3 DNA-binding activity (80 μg/ml: 21.37% inhibition).

Conclusions: Anthriscus sylvestris exerts anti-colon cancer effects by targeting STAT3 signaling, supported by network pharmacology and experimental validation. Its petroleum ether extract demonstrates significant therapeutic potential, providing a scientific foundation for developing AS-based anti-cancer foods or drugs.