Abstract: Objective: Network pharmacology, molecular docking technology, and CCK8, wound healing, cell invasion, plate colony formation assay were used to explore the effect of apigenin in the treatment of gastric cancer. Methods: The ChEMBL, UniProt, and GeneCards databases and jvenn tools were used to obtain the potential anticancer targets of apigenin. Then, the PPI network analysis, gene ontology (GO) enrichment analysis and Kyoto Encyclopedia of genes and genomes (KEGG) pathway enrichment analysis of key targets were used to investigate the mechanism, and Auto dock software was used for further molecular docking. Lastly, the inhibitory effect of apigenin on gastric cancer cells was verified through cell counting kit-8 (CCK8) experiments. Results: There were 52 potential targets of apigenin in the treatment of gastric cancer. GO analysis showed that apigenin might exert its anti-gastric cancer mechanism by regulating signal transduction, chemical synaptic transmission, and proteolysis. KEGG analysis showed that apigenin played a role by regulating the neural active ligand receptor interaction, nitrogen metabolism and other signaling pathways. The molecular docking results showed that the binding energy of ligand and receptor was: AChE|−9.1|>MAOA|−8.8|>CA2|−7.4|>DPP4|−7.3|>CA1|−7.2|=GRM5|−7.2|>ADA|−6.8|>CASP3|−5.8|, indicating that the ligand and receptor had good binding affinity. In vitro tests exhibited that apigenin possessed significant inhibitory effect on gastric cancer cell lines, which might be due to its ability to reducing the expression of MAOA, DPP4, AChE and increasing the expression of CA2. Conclusion: Apigenin has the effect of anti-gastric cancer, and the main targets involved in its effect are AChE, CA1, CA2, CASP3, ADA, MAOA, DPP4, GRM5, which shows the advantages of apigenin in multi-component, multi-target, and multi-channel, and provides a reference for the follow-up study of the anti-gastric cancer effect of apigenin.