Pancreatic cancer is a highly lethal cancer with limited treatment options. The number of pancreatic cancer patients is increasing rapidly worldwide. Many natural products have been shown to have anticancer activity in a range of studies. Sotetsuflavone is derived from Cycas revoluta Thunb. and exhibits anticancer activity. The present study incorporates network pharmacology, single gene survival analysis, gene expression analysis and molecular docking to reveal the mechanism of Sotetsuflavone in the treatment of pancreatic cancer. Initially, it was evaluated through multiple databases for a comprehensive pharmacological evaluation of Sotetsuflavone. Then, the target information of Sotetsuflavone and pancreatic cancer was searched and screened using public databases. According to the results of matching the potential targets of Sotetsuflavone with the targets of pancreatic cancer, the protein-protein interaction (PPI) network was constructed by using the STRING database and imported into Cytoscape 3.9.0 for the network analysis to screen the hub targets, and then classify and co-expression analysis was carried out on the hub targets. Then, we conducted Gene Ontology (GO) analysis and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway enrichment analyses of potential and hub targets and discovered that these targets were involved in pancreatic cancer-related signalling pathways. In addition, the GEPIA 2 platform was used to be performed for single gene survival analysis and gene expression analysis, and AutoDock Vina was employed for molecular docking analysis. In total, we have acquired 31 hub targets for the treatment of pancreatic cancer by Sotetsuflavone, namely ABCB1, AURKA, CDK1, and so on. Kaplan-Meier survival analyses demonstrated that ABCB1, AURKA, CDK1, HDAC6, MET, and MMP3 are promising hub targets that can be used as biomarkers for pancreatic cancer diagnosis and prognosis. These hub targets are highly expressed in pancreatic cancer tissues compared to normal tissues. The molecular docking results showed a strong binding capacity of Sotetsuflavone to these hub targets. In summary, it is proposed that Sotetsuflavone is a new anticancer drug, which can regulate cancer-related signalling pathways such as pancreatic cancer by inhibiting the activities of ABCB1, AURKA, CDK1, HDAC6, MET, and MMP3, which are hub targets with up-regulated expression in pancreatic cancer tissues, in order to treat pancreatic cancer. However, it also requires a series of in vivo and in vitro studies to ensure its safety and efficacy.