Intervention Effect of Betulinic Acid on Breast Cancer and Core Pathway Prediction
Objective:To explore the potential mechanism of betulinic acid in the treatment of breast cancer.Methods:Drug and disease targets were obtained through PharmMapper,Genecards,Drugbank and Therapeutic Target Database(TTD).The protein-protein interaction was re-vealed in the String.Metascape was used for Gene Ontology(GO)and Kyoto Encyclopedia of Genes and Genomes(KEGG)enrichment a-nalysis.Molecular docking was carried out by Autodockvina and visual analysis was carried out by Pymol.Kaplan-MeierPlotter was employed to analyze the prognosis and survival of the key targets.The effect of betulinic acid on the proliferation and migration of breast cancer cells was detected by MTT method,clone formation assay,wound healing assay and Transwell assay.AO/EB staining and Annexin V-FITC/PI stai-ning were performed to observe the effect of betulinic acid on cell apoptosis,while its effect on PI3K/AKT/mTOR signaling pathway was de-tected by Western blot.Results:From the protein-protein interaction network of common targets of betulinic acid and breast cancer,33 nodes and 100 edges were selected,with an average node degree of 6.06.The core targets selected were NR3C1,SRC,ALB,MAPK8,PGR and GRB2.The binding energy of betulinic acid with the targets were all less than 5 kcal/mol.There were 473 entries observed in the biologi-cal process of GO function enrichment,mainly including cellular response to hormone stimulus,intracellular receptor signaling pathway,and hormone-mediated signaling pathway,42 entries in molecular function,mainly including lipid binding,protein kinase activity,and transcription factor binding,and 7 entries in cellular components,mainly involving receptor complexes,transcription factor complexes,membrane raft,etc.A total of 172 signaling pathways were obtained from KEGG enrichment analysis,including PI3K/AKT,Ras,ErbB,GnRH and EGFR signaling pathways.In vitro experiments showed that betulinic acid inhibited the proliferation of breast cancer cells in a dose-and time-dependent man-ner,as well as the invasion and migration of the cells.Betulinic acid promoted the apoptosis qualitatively and quantitatively and down-regulate the protein expressions of PI3K,AKT and mTOR(P＜0.01).Conclusion:Betulinic acid can inhibit the proliferation,migration,apoptosis and other malignant biological behaviors of breast cancer cells MCF-7,and its mechanism may be related to the inhibition of the proteins in PI3K/AKT/mTOR signaling pathway.
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