Network pharmacology‑based analysis and experimental verification of the inhibitory role of luteoloside on gastric cancer cells via the p53/p21 pathway.

The present study aimed to investigate the inhibitory effect of luteoloside on the proliferation, migration and invasion of gastric cancer (GC) cells based on network pharmacology and experiments. GC-associated targets were obtained from the GeneCards and Online Mendelian Inheritance in Man databases. Gene Ontology functional enrichment analysis and Kyoto Encyclopedia of Genes and Genomes pathway enrichment analysis were performed using the Database for Annotation, Visualization and Integrated Discovery. Protein-protein interaction (PPI) networks and herb-active ingredient-target gene-signaling pathway networks were constructed using the Search Tool for the Retrieval of Interacting Genes and proteins and Cytoscape software to analyze core target genes and pathways. In addition, the alkaline comet assay was performed to assess DNA damage, demonstrating that luteoloside induces DNA double-strand breaks in a concentration-dependent manner, as indicated by increased comet tail lengths. γ-H2AX detection through western blot analysis further corroborated these findings, showing significant upregulation of this DNA damage marker in luteoloside-treated GC cells. The human GC cell line NCI-N87 was utilized for experiments to investigate the impact of different doses of luteoloside on cell proliferation, invasion and migration using Cell Counting Kit-8, scratch-wound and Transwell assays, respectively. The underlying molecular mechanism of luteoloside was explored using western blot analysis. The successfully constructed PPI network revealed the p53, Akt1, Bcl-2 and Caspase-3 proteins as the core targets, all of which showed good binding activity with luteoloside. The experiments demonstrated that luteoloside treatment significantly inhibited GC-cell proliferation, migration and invasion. The western blot results showed notable concentration-dependent upregulation of p53 and p21 protein expression and downregulation of Bcl-2 protein expression following luteoloside treatment. Overall, luteoloside inhibited the proliferation, migration and invasion of GC cells by activating the p53/p21 signaling pathway.