OBJECTIVE  To integrate network pharmacology analysis and in vitro cell experiments to study the mechanism of action of sempervirine(SPV) in the treatment of nasopharyngeal carcinoma(NPC).

METHODS  PhamMapper, SwissTargetPrediction and other databases were used to collect the targets of SPV, and the target genes related to NPC were obtained from Gene card, OMIM and other databases. By constructing the Venn diagram, the common targets of drugs and diseases were obtained. The STRING online platform was used to construct the protein interaction network of common targets, and the core targets were obtained by topological parameter screening of the network by Cytoscape 3.9.1 software, the binding activity and stability of SPV and the core target were verified by molecular docking and molecular dynamics simulation. The effect of SPV on the proliferation ability of NPC cells(HK-1) was detected by CCK-8 assay. The Transwell assay, cell scratch assay, cell adhesion assay was used to observe the effect of SPV on the migration and invasion ability of HK-1 cells. Western blotting was used to intervene in the protein expression of EGFR and p-EGFR in NPC cells HK-1 after intervention with SPV.

RESULTS  A total of 3565 SPV targets and 182 target genes related to NPC were screened through network pharmacology, and a total of 91 potential targets of SPV in the treatment of NPC were screened after intersection, and KEGG analysis was significantly enriched in PI3K-Akt, Ras and other pathways, and showed that gene function was related to protein phosphorylation. The core targets such as EGFR, SRC, ALB, ESR1, CASP3 and MMP2 were screened by Cytoscape 3.9.1, and molecular docking experiments showed that SPV had good binding activity with multiple core targets. Furthermore, molecular dynamics simulation experiments proved that SPV had good stability in binding to EGFR. Cell experiments showed that compared with the control group, SPV could effectively inhibit the proliferation, migration and invasion of HK-1 cells, and down-regulated the level of p-EGFR/EGFR protein.

CONCLUSION  SPV may inhibit the proliferation and invasion of NPC cells by regulating EGFR.