Objective To explore the mechanism of cordycepin (Cor) in the treatment of non-small cell lung cancer ( NSCLC ) by network pharmacology, molecular docking and in vitro experiments. Methods Multiple databases were used to search the targets of Cor and the therapeutic targets of NSCLC, and the common targets were screened out. STRING database and Cytostape were used to analyze and visualize the protein-protein interaction networks (PPI). GO and KEGG enrichment analysis was performed using the DAVID database, and the results were visualized using the online tool “Weishengxin”. AutoDock Vina was used to verify the molecular docking of Cor with the core target, and the binding energy was calculated. PyMol was used to visualize the results. MTT assay was used to detect the effect of Cor on the proliferation of NSCLC A549 cells. Western blot was used to detect the effect of Cor on the expression of core target proteins PARP1, P53 and AKT1. Results A total of 215 targets of Cor and 1238 therapeutic targets of NSCLC were screened, and 122 common targets were obtained. After PPI analysis, the core targets PARP1, P53 and AKT1 were selected according to Degree value for verification. The results of GO and KEGG enrichment analysis showed that biological process (BP) mainly involved apoptosis process, positive and negative regulation of gene expression, cellular component ( CC ) mainly involved cytoplasm and nucleus, molecular function (MF) mainly involved protein binding and enzyme binding, and signaling pathways mainly involved cancer, PI3K / AKT and other signaling pathways. Molecular docking showed that the three targets of PARP1, P53 and AKT1 were highly correlated with the treatment of NSCLC. In vitro experiments showed that Cor could inhibit the proliferation of human NSCLC A549 cells, significantly up-regulate the expression of P53 and PARP1 and down-regulate the expression of AKT1. Conclusion Cor inhibits the proliferation of human NSCLC A549 cells, and the mechanism may be related to the regulation of PARP1, P53 and AKT1 expression.