Network pharmacology,molecular docking and experimental validation were employed to explore the potential therapeutic targets and mechanisms of scutellarin (SCU) in the treatment of breast cancer.The targets of SCU were predicted through SwissTargetPrediction database and TargetNet database,and the potentical anti-breast cancer targets of SCU were obtained by mapping them with the potential targets associated with breast cancer retrieved from GeneCards,OMIM,TTD databases.The STRING database was applied to construt the protein-protein interaction networks and topological analysis was performed by Cytoscape 3.7.2 software.GO functional enrichment analysis and KEGG pathway enrichment analysis of intersecting targets was carried out using the DAVID database.The building of SCU to key targets was validated through molecular docking study using Autodock Vina software.The expression of relevant target genes in Clinical case samples obtained from the Cancer Genome Atlas (TCGA) database were analyzed.At last,CCK-8 assay was performed to explore the anti-proliferative effect of SCU under different concentrations and the effects of SCU on the protein expression of TNF-α,EGFR and HIF-1α signaling pathway in human breast adenocarcinoma MCF-7 cells were examined by Western blot.As a result,a total of 90 potential targets were screened out,and five key targets were obtained after topological analysis,among which EGFR,TNF,CASP3,PTGS2,and MAPK14 are closely related to the anti-breast cancer effect of SCU.Multiple signaling pathways such as C-type lectin receptor signaling pathway,Apoptosis,Human cytomegalovirus infection,HIF-1 signaling pathway and IL-17 signaling pathway were involved in the anti-breast cancer effect of SCU.Molecular docking results showed that SCU has an excellent binding effect with key target proteins EGFR and TNF.The results of clinical samples revealed that the key target genes were significantly changed in breast caner patients compared with the healthy population.In vitro cell experiments showed that SCU (40-160 μmol/L) significantly inhibited the proliferation of human breast adenocarcinoma MCF-7 cells in a dose-dependent manner (P < 0.01 or P < 0.05).Further western blot assay confirmed SCU up-regulated EGFR protein expression and down-regulated TNF-α  and HIF-1 α  protein expression.In conclusion,SCU exerts anti-cancer effect through multiple targets and signal pathways.This study provided a theoretical basis and reference for the exploration of SCU on clinical research and product development.