Exploring the molecular mechanism of Coptis-cinnamon in combating gastric cancer via the MAPK Pathway based on network pharmacology.

Gastric cancer (GC) ranks as the fifth most common malignancy worldwide. Current treatments are limited by side effects and drug resistance, highlighting the need for novel therapies. Traditional Chinese medicine (TCM) pair Coptis-Cinnamon (HL-RG) shows promise against GC, but its mechanism remains unclear. Active components and targets of HL-RG were obtained from TCMSP, and GC-related genes from GEO and GeneCards. Intersection targets were analyzed via STRING for PPI network construction. GO and KEGG enrichment analyses were performed using DAVID to identify key pathways and hub genes. Clinical relevance, mutations, immune infiltration, and drug sensitivity were analyzed. Molecular docking validated interactions between core components and hub genes. In vitro experiments (CCK-8, flow cytometry, Transwell, qRT-PCR, Western blot) were conducted in AGS and HGC-27 cells to verify anti-GC effects. To verify the causal relationship, we conducted rescue experiments using the MEK inhibitor U0166 and the MAPK agonist EGF. Sixteen active components and 499 targets of HL-RG were identified, with 55 common targets screened from 3,194 GC-related DEGs. Enrichment analyses revealed involvement in inflammatory responses and the MAPK pathway, identifying PDGFRB, EGFR, MMP2, MMP9, and KIT as hub genes. Molecular docking showed binding affinities between core components (berberine, berberrubine) and hub genes. In vitro experiments confirmed that HL-RG inhibits GC cell proliferation, induces apoptosis, and suppresses migration and invasion via MAPK pathway regulation. Rescue experiments demonstrated that U0126 phenocopied HL-RG effects, while EGF significantly reversed HL-RG-induced functional changes and p-ERK reduction, confirming that HL-RG acts through the MAPK/ERK pathway. HL-RG exerts anti-GC effects by targeting the MAPK pathway and its hub genes, providing a scientific basis for its clinical application.