Exploring effects and potential mechanisms of cryptotanshinone on A2780/DDP cisplatin-resistant cells based on transcriptome sequencing technology
Objective To investigated the effects of cryptotanshinone(CPT)combined with cisplatin(DDP)on A2780/DDP cells and its potential mechanism of action based on transcriptome sequencing technology.Methods The A2780 and A2780/DDP cells were cultured in vitro,and the cells were divided into control group,DDP group,and DDP+CPT experimental group.MTS cell proliferation and cytotoxicity detection Kit were used to detect the cell survival rate,scratch assay was used to detect the migratory ability,and Transwell assay was used to detect the migratory and invasive ability,to investigate the effect of DDP+CPT intervention on A2780/DDP cells.The mRNA of the above grouped cells was obtained for transcriptome sequencing,screening for differentially expressed genes,topology analysis was used to obtain the core target of CPT to improve the resistance of A2780/DDP to DDP,the gene ontology(GO)function and the Kyoto encyclopedia of genes and genomes(KEGG)enrichment analysis were conducted for differentially expressed genes,molecular docking technology was used to study the binding ability of CPT to the core target and real-time reverse transcription quantitative polymerase chain reaction(RT-qPCR)was used to detect the mRNA expression of the core target.Results The resistance index of A2780/DDP cells was four,DDP+CPT intervention significantly inhibited the cell viability,cell migration and cell invasion ability of A2780/DDP-resistant strains(P＜0.001).The results of differentially expressed genes screening showed that there were 253 differentially expressed genes in the drug group as compared with the model group,among which JUN proto-oncogene gene(JUN),Toll-like receptor 2(TLR2),quinone oxidoreductase 1(NQO1),and nitric oxide synthase 2(NOS2)were significantly up-regulated(P＜0.05),while chemokine receptor 4(CXCR4)was significantly decreased(P＜0.05).The results of GO and KEGG suggested that CPT might affect the TLR signaling pathway,B-cell receptor signaling pathway,and interleukin-17 signaling pathway and other mechanisms,thereby improving the drug resistance of A2780/DDP cells to DDP.RT-qPCR results showed that CPT up-regulated the mRNA expression of JUN,TLR2,NOS2,NQO1(P＜0.001),inhibited CXCR4 mRNA expression(P＜0.001)to improve the drug resistance of A2780/DDP to DDP.Conclusion CPT could improve the resistance of A2780/DDP-resistant cells to DDP,and the mechanism may be related to the up-regulation of the mRNA expression of JUN,TLR2,NOS2,NQO1 and the down-regulation of CXCR4 mRNA expression by CPT.
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