Objective To investigate the anti-gastric cancer effects of silybin and its underlying molecular mechanisms based on network pharmacology and experimental validation.

Methods Potential targets of silybin were retrieved from the PubChem, SuperPRED, and SwissTargetPrediction databases, while gastric cancer-related targets were screened from the GeneCards, OMIM, and DrugBank databases. The intersecting targets of silybin and gastric cancer were imported into the STRING database to construct the protein-protein interaction (PPI) network, followed by Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) enrichment analyses. The CCK-8 assay was used to evaluate the effects of silybin on the viability of AGS and HGC27 cells. Colony formation, wound healing, and transwell migration assays were performed to assess the effects of silybin on the proliferation and migration of AGS cells, respectively. Western blotting was employed to analyze the expression of proteins associated with the phosphatidylinositol 3-kinase/protein kinase B (PI3K/AKT) signaling pathway.

Results Network pharmacology analysis identified 92 intersecting targets between silybin and gastric cancer. KEGG enrichment analysis revealed significant enrichment of the PI3K/AKT signaling pathway. Experimental validation showed that the IC₅₀ values of silybin in AGS and HGC27 cells were 87.33 μmol/L and 87.49 μmol/L, respectively. Furthermore, silybin dose-dependently inhibited the proliferation, colony formation, and migration of AGS cells (P < 0.05). Western blotting results demonstrated that silybin dose-dependently decreased the protein expression levels of phosphorylated PI3K (p-PI3K), phosphorylated AKT (p-AKT), and matrix metalloproteinase 9 (MMP9), while upregulating tumor protein P53 (TP53) (P < 0.05). Notably, insulin-like growth factor-1 (IGF-1) partially reversed these effects of silybin (P < 0.05).

Conclusion Silybin suppresses gastric cancer cell proliferation and migration by suppressing the phosphorylation activation of the PI3K/AKT signaling pathway, downregulating MMP9 expression, and upregulating TP53 expression.

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