Exploring the potential mechanism of artemisinin against gastric cancer based on network pharmacology, machine learning and experimental validation

Background Artemisinin, a natural active component extracted from Artemisia annua , exhibits potential anti-gastric cancer activity; however, its specific targets and molecular mechanisms remain unclear. Methods Network pharmacology was used to predict the potential targets of artemisinin. Differential expression analysis and weighted gene co-expression network analysis (WGCNA) were performed to identify gastric cancer-related genes. Candidate targets of artemisinin against gastric cancer were obtained by intersecting the above datasets. Three machine learning algorithms, namely random forest, LASSO regression, and SVM-RFE, were further employed to screen hub targets, followed by molecular docking to evaluate the binding affinity between artemisinin and the hub targets. Finally, anti-gastric cancer effects and mechanisms were validated by in vitro experiments (CCK-8, EdU, colony formation, Transwell and Western blot). Results A total of 27 candidate targets of artemisinin against gastric cancer were identified. Cross-screening by the three machine learning algorithms ultimately identified TOP2A , CCNB2 , and CCNA2 as hub targets. Molecular docking revealed low binding energies (all below −5.0 kcal/mol) between artemisinin and these three targets, suggesting potential binding affinity. In vitro experiments demonstrated that artemisinin significantly downregulated the mRNA expression levels of the three targets and suppressed the proliferation, migration, and invasion of gastric cancer cells. Mechanistic investigation showed that artemisinin treatment decreased p-ERK levels, and the anti-gastric cancer effects of artemisinin were partially reversed by the ERK pathway activator C16-PAF. Conclusions In vitro , artemisinin suppresses ERK/MAPK pathway activation and reduces the mRNA expression of cell cycle-related TOP2A , CCNB2 and CCNA2 , leading to inhibited proliferation, migration and invasion of gastric cancer cells.