Mechanism of dioscin in reversing doxorubicin resistance of breast cancer based upon cell assays and net-work pharmacology
OBJECTIVE To explore the effect of dioscin(DIO)on doxorubicin(DOX)resistance in human breast cancer MCF-7/DOX cell and elucidate its possible mechanism.METHODS MTT assay was utilized for detecting the viability of MCF-7/DOX cell after treating with different concentrations of DIO.MCF-7/DOX cell was exposed to non-toxic concentrations of DIO plus DOX for observing the reversal effect of DIO on DOX resistance by methylthiazolyldiphenyl tetrazolium(MTT),West-em blot and quantitative real-time polymerase chain reaction(qRT-PCR).Network pharmacology was utilized for predicting the potential targets of DIO against DOX resistance in breast cancer.Protein-protein interaction(PPI)network was constructed through a STRING database and Cytoscape 3.9.1 software for selecting core targets.Gene Ontology(GO)and Kyoto Encyclo-pedia of Genes and Genomes(KEGG)pathway enrichment analyses were further performed through the David database.Finally qRT-PCR was performed to verify the mRNA expression of core targets.RESULTS At a concentration of 0.375-1.56 μg·mL-1,DIO displayed little cellular cytotoxicity.After exposure to DIO 0.75 μg·mL-1 plus different concentrations of DOX,half maxi-mal inhibitory concentration(IC50)of DOX declined with a reversing multitude of 2.02 and protein/mRNA expressions of MDR1/MRP1 became down-regulated(P＜0.05).It implied that DIO reversed DOX resistance.Network pharmacology prediction yielded 78 potential targets for DIO reversing DOX resistance in breast cancer.Some core targets included protein kinase B(AKT1),tumor protein P53(TP53),apoptosis regulator BCL2(BCL2),interleukin-6(IL-6),caspase 3(CASP3)and jun proto-oncogene(JUN)involved in lipid and atherosclerosis,neoplastic pathways and other signal pathways.0.75 μg·mL-1 DIO plus DOX up-regulated mRNA expressions of TP53 and CASP3(vs.control group,P＜0.05)and down-regulated mRNA expressions of AKT1 and IL-6(vs.DOX group,P＜0.05).With a tendency of down-regulating BCL2 mRNA,it had no obvi-ous effect on JUN mRNA level.CONCLUSION DIO may reverse DOX resistance in breast cancer through modulating the core targets of AKT1,TP53,BCL2,IL-6 and CASP3.
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