Objective To explore the anti-tumor effect and related mechanisms of Toosendanin on human nasopharyngeal carcinoma CNE2 cells based on network pharmacology. Methods The molecular regulatory pathways of melanoidin acting on nasopharyngeal carcinoma were screened out by network pharmacology. Human nasopharyngeal carcinoma CNE2 cells were cultured in vitro, and the effects of different melanoidin concentrations on the proliferation of CNE2 cells were detected at different time points (0, 24, 48, 72 h) using the CCK-8 method. Western blot experiments verified the mechanism of network pharmacological screening; The scratch assay, Transwell assay and flow cytometry assay were used to evaluate the effects of toonein on the migration, invasion, apoptosis and cell cycle of CNE2 cells. The Western blot experiment was used to detect changes of key proteins Bcl -2, Bax and Caspase-3 in the apoptotic pathway.Results Based on the enrichment of GO and KEGG in network pharmacology, it was found that melanoidin might exert its effect through the PI3K/op-AKT pathway. Western blot experiments confirmed that melanoidin could reduce the protein levels of phosphorylated PI3K(P op-PI3K) and phosphorylated AKT (P op-Akt), while the expressions of PI3K and AKT showed no significant changes. Compared with the control group, tooneein could effectively inhibit the proliferation of CNE2 cells in a time and concentration-dependent manner. The concentration groups of 0.08 μmol/L, 0.16 μmol/L, and 0.32 μmol/L were selected for the subsequent experiments. Tooneein significantly inhibits the migration and invasion of CNE2 cells, simultaneously induces their apoptosis, and causes the cell cycle to stagnate at the S phase. And it led to an increase in the expression of Bax and Caspase 3 in CNE2 cells and a decrease in the expression of Bcl 2 protein (all data differences were statistically significant and P<0.05). Conclusion Toosendanin efficiently inhibits the proliferation and migration of human nasopharyngeal carcinoma CNE2 cells, induces apoptosis, and alters the cell cycle. Its mechanisms of action may be related to the apoptosis pathway and the PI3K-AKT signaling pathway.These findings provide a theoretical basis for the application of Toosendanin in the treatment of nasopharyngeal carcinoma