Non-small cell lung cancer (NSCLC) is the most prevalent form of lung cancer. However, its mechanism and therapeutic strategy remain to be clarified. Mangiferin (MGF) is a flavonoid derived from the leaves of mango trees that has many pharmacological and physiological effects. The aim of this research is to elucidate the mechanism of MGF in treating NSCLC using network pharmacology, molecular docking, and the biological effect of MGF in NSCLS cell line. The targets of MGF were screened by the TCMSP, Swiss Target Prediction, and SuperPred databases. The targets related to MGF were obtained from the GeneCards, OMIM, PharmGkb, TTD, DrugBank, and DisGeNET databases, and the intersected targets were obtained by Venny2.1.0. The protein-protein interaction (PPI) network was drawn through the String database and Cytoscape software. Next, the pathway enrichment analyses of Kyoto Encyclopedia of Genes and Genomes (KEGG)

enrichment analyses were performed by the Bioconductor platform. Finally, molecular docking was conducted using Autodock Vina. The result showed that 74 potential targets of MGF were chosen and 53 overlapping targets were obtained. In the PPI network, HSP90AA1, ESR1, PTGS2, CXCR4, and TLR4 were the core targets. KEGG analyses showed that the main pathways of MGF in treating NSCLC involved PI3K-Akt, NF-kappa B, and MAPK signaling pathways. In molecular docking, MGF had a good affinity with the core targets. In vitro experiments, MGF was shown to significantly restrain the proliferation of NSCLC cells (A549, NCI-H460, and NCI-H1299 cells) in a dose- and time-dependent manner. In conclusion, MGF can treat non-small cell lung cancer through multiple targets and pathways which could provide new ideas for further research on the protective mechanism and clinical application of MGF against NSCLC.