A Network Pharmacology-based Study On The Effect Of Angelicin In The Treatment Of Osteosarcoma And Its Mechanism

Objective:To investigate whether angelicin has antitumor effects on osteosarcoma and its mechanism.Methods:Using network pharmacology and molecular docking,we screened the potential therapeutic targets of Angelicin for osteosarcoma.We used this as a basis to validate the effects of Angelicin on osteosarcoma through in vitro experiments.First,by applying Venn diagrams to map the targets of Angelicin to the targets of osteosarcoma,the targets in the intersection part were considered potential therapeutic targets.Second,the String database and Cytoscape software were used to analyze potential therapeutic targets,and protein-protein interaction(PPI)networks were constructed and analyzed to obtain hub genes.Gene Ontology(GO)and Kyoto Encyclopedia of Genes and Genomes(KEGG)functional enrichment analyses of potential therapeutic targets were performed using R software.Then,molecular docking of Angelicin and pivotal targets was performed using Autodock Tool software as well as Autodock vina software,and the results were visualized using Pymol software.Finally,based on the results of the network pharmacological analysis,in vitro experiments were applied to verify the effects of Angelicin on osteosarcoma MG63 cells,such as proliferation,apoptosis,and migration,as well as to detect the effects of Angelicin on the m RNA expression of hub genes in osteosarcoma MG63 cells using RT-PCR technique.Results:Venn diagrams showed that 19 of 33 Angelicin-related targets were potential therapeutic targets for Angelicin in treating osteosarcoma.By constructing and analyzing PPI,Degree≥2,Closeness Centrality(CC)≥0,and Betweenness Centrality(BC)≤0.714286 were used as screening conditions,and a total of four hub targets,BAX,BCL-2,BIRC 2,and CASP9,were screened.Molecular docking results showed that these four hub targets were well connected to Angelicin(<-4kcal/mol).Functional enrichment analysis of potential therapeutic targets of Angelicin showed that these targets act through multiple signaling pathways,among which apoptosis-multiple species,small cell lung cancer,NF-κB signaling pathway,apoptosis,and p53 signaling pathway were dominant.The CCK8 assay showed that Angelicin inhibited the proliferation of MG63 cells,and the wound healing assay showed that Angelicin inhibited MG63 cell migration activity.RT-PCR results showed that Angelicin up-regulated the expression of anti-apoptotic genes BCL-2 and CASP9 genes and down-regulated the expression of pro-apoptotic BAX and BIRC2 genes.Conclusions:Angelicin can affect the expression of four hub targets,BAX,BCL-2,BIRC 2,and CASP9,which in turn affect the signaling network where the hub targets are located,producing an effect of inhibiting the proliferation and migration of osteosarcoma MG63 cells and promoting apoptosis of osteosarcoma MG63 cells.In conclusion,the use of Angelicin in treating osteosarcoma is expected to be a new idea in treating osteosarcoma in clinical medicine.