Abstract: Objective:To predict the related targets and pathways of Alisol B on nasopharyngeal carcinoma(NPC) using network pharmacology analysis, and to explore its mechanism of action and validate it through experiments.Methods:The Pubchem, Pharmmapper, GeneCards and other databases were searched to obtain the related targets of Alisol B on NPC and Epstein-Barr virus(EBV), and the intersection targets between them were obtained.Then a "Drug-Target-Disease-Pathway" network was constructed using Cytoscape software.Finally, Gene Ontology (GO) functional enrichment analysis and Kyoto Encyclopedia of Genes and Genomes (KEGG)pathway enrichment analysis were performed using the David database.The proliferation inhibitory effect of Alisol B on CNE-2 cells of NPC at different concentrations was detected by CCK-8 and plate cloning experiments.The expression of predicted pathway target proteins was verified by western blotting.Results:Network pharmacology analysis identified 71 potential targets of Alisol B on NPC, and the 10 most relevant targets were screened out according to the degree value from high to low.GO annotation yielded 544 molecular function entries, and the top 15 significantly enriched entries were obtained before ranking the results in ascending order of P value.KEGG analysis showed that it was closely related to PI3K-Akt signaling pathway; the key genes involved included: Akt1, MAPK1, HSP90AA1, MAPK14, etc.The results of CCK-8 and plate cloning experiments showed that Alisol B could effectively inhibit the proliferation of NPC cells, and the inhibitory effect was enhanced with increasing drug duration and concentration.Western blotting confirmed that Alisol B significantly down-regulated the expression levels of Akt and PI3K proteins(P< 0.05).Conclusion:Alisol B can effectively inhibit the proliferation of NPC CNE-2 cells, and its anti-NPC mechanism may be related to the inhibition of PI3K-Akt signaling pathway activity.