Objective To explore the mechanism of morin, an active ingredient of Sangzhi (Mori Ramulus), against gastric cancer based on network pharmacology, molecular docking and in vitro experiments. Methods Potential targets of Mori Ramulus and gastric cancer-related targets were screened by network pharmacology, and the intersection of targets was obtained. Protein-protein interaction (PPI) network topological analysis, gene ontology (GO) and Kyoto encyclopedia of genes and genomes (KEGG) enrichment analysis, and GEO dataset validation were used to screen core targets. Molecular docking and molecular dynamics simulation were applied to verify the binding ability of morin (the active ingredient of Mori Ramulus) to phosphatidylinositol 3-kinase regulatory subunit 1 (PIK3R1) and the stability of the formed complex. In in vitro experiments, human gastric adenocarcinoma AGS cells were treated with 100, 200, 300, 400 μmol/L morin alone or combined with phosphatidylinositol 3-kinase (PI3K) agonist 740Y-P. Cell proliferation ability was detected by CCK-8 assay and plate clone formation assay. Cell apoptosis rate and cell cycle distribution were detected by flow cytometry. Expressions of PI3K/protein kinase B (Akt) pathway, apoptosis and cycle related proteins were detected by Western blotting. Results A total of 178 potential targets of Mori Ramulus, 13 100 gastric cancer-related targets, and 159 intersecting targets were identified. Enrichment analysis showed that the key pathway was the PI3K/Akt pathway. Morin had strong binding affinity with PIK3R1, was significantly highly expressed in gastric cancer tissues, and the constructed morin-PIK3R1 complex maintained excellent stability. The in vitro experiment results showed that morin could inhibit AGS cells proliferation in a dose-dependent manner, induce G₀/G₁ phase arrest, and promote cell apoptosis (P < 0.05, 0.01). Meanwhile, morin downregulated the expressions of PI3K/Akt pathway, B-cell lymphoma-2 (Bcl-2), cyclin D1 (CCND1), cyclin-dependent kinase 4 (CDK4) and cyclin-dependent kinase 6 (CDK6) protein (P < 0.05, 0.01), and upregulated the expressions of Bcl-2 associated X protein (Bax) and p21 (P < 0.01). Combined treatment with 740Y-P significantly reversed the effects of morin on proliferation, apoptosis and cell cycle (P < 0.05, 0.01). Conclusion Morin can exert anti-gastric cancer activity by targeting and stably binding to PIK3R1, inhibiting the activation of PI3K/Akt pathway, inducing cell cycle arrest, promoting apoptosis, and inhibiting proliferation in gastric cancer cells.

morin  /  gastric cancer  /  bioinformatics  /  PI3K/Akt pathway  /  cell apoptosis  /  cell proliferation