Role and mechanism of gambogic acid in inhibiting bladder cancer based on network pharmacology and experimental verification
Objective To determine the role and mechanism of gambogic acid(GA)in inhibiting bladder cancer based on network pharmacology and experimental verification.Methods PubChem,PharmMapper,GeneCards databases and Venny 2.1.0 were retrieved to obtain the potential target genes of GA in inhibiting bladder cancer.Protein-protein interaction diagram was made by STRING database and Cytoscape 3.9.1 software.The DAVID database was used for GO and KEGG enrichment analysis and visualization.CCK-8 and colony formation assays were used to detect the effect of GA on the proliferation of UM-UC-3 cells.Wound healing assay and Transwell assay were used to detect the effect of GA on the migration and invasion of UM-UC-3 cells.Flow cytometry was used to detect the effect of GA on the apoptosis of UM-UC-3 cells.Western blot was used to detect the effect of GA on the protein expression levels of PI3K,p-PI3K,AKT,p-AKT and FoxO1 in UM-UC-3 cells.Results Totally 57 potential targets of GA in inhibiting bladder cancer were screened,and the core targets was EP300.GO analysis showed that the inhibitory effect of GA on bladder cancer may involved 26 biological processes,17 cellular components,and 28 molecular functions.KEGG pathway enrichment analysis showed that the inhibitory effect of GA on bladder cancer might be related to PI3K/AKT,FoxO1 and HIF-1 signaling pathways.Cytological experiments showed that after 24 h GA treatment,the proliferation,migration and invasion of UM-UC-3 cells were decreased,and the apoptosis rate was increased as compared with those of the control group.The protein expression levels of PI3K,p-PI3K,and p-AKT in UM-UC-3 cells were down-regulated,and the protein expression level of FoxO1 was up-regulated.Conclusion GA inhibits the proliferation,migration and invasion of UM-UC-3 cells,and promotes the apoptosis of UM-UC-3 cells,whose mechanism may be related to the PI3K/AKT/FoxO1 signaling pathway.
network pharmacologygambogic acidbladder cancerproliferationmigration and invasionapoptosis
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