Objective To investigate the reversal effect and mechanism of cinobufagin(CBG)on cisplatin resist-ance in human ovarian cancer cells.Methods A2780 cell line and its cisplatin-resistant cell line A2780/DDP are common ovarian cancer cells in clinic,so these two cell lines were selected as the research objects.The cell viabil-ity was detected by cell Counting Kit-8(CCK-8)assay,and the cell proliferation ability was detected by plate clo-ning and 5-ethynyl-2′-deoxyuridine(EdU)assay.Hoechst staining was used to observe cell apoptosis.Cell scratch test and Transwell test were used to evaluate cell migration and invasion ability.Western blot and quantitative reverse transcription PCR(RT-qPCR)were used to detect the protein and mRNA expressions of phosphatidylinosi-tol 3-kinase/protein kinase(PI3K/AKT)signaling pathway and epithelial-mesenchymal transition(EMT).Re-sults Compared with A2780 cells,the drug resistance indexes of A2780/DDP cells were 5.636,5.864,5.695,respectively.After treatment of A2780/DDP cells with CBG(2,4,6 mg/ml),the reversal resistance indexes were 1.617,2.570,3.461,respectively.CBG treatment significantly increased the level of apoptosis and inhibi-ted the proliferation,migration and invasion of the cells in a concentration-dependent manner(P<0.05).Western blot results showed that compared with A2780 cells,the relative ratio of P-PI3K/PI3K and P-AKT/AKT protein levels,as well as the protein expression of N-cadherin,Vimentin,and Snail were higher in the control group(A2780/DDP)cells,while the protein expression of E-cadherin was lower(tP-PI3K/PI3K=8.115,tP-AKT/AKT=17.62,tN-cadherin=6.126,tVimentin=4.001,tSnail=17.333,tE-cadherin=4.620,P<0.01);As the dose of CBG increased,the protein expression levels of P-PI3K,P-AKT,N-cadherin,Vimentin,and Snail in drug-resistant cells de-creased,while the protein expression level of E-cadherin increased(FP-PI3K=268.5,FP-AKT=190.5,FN-cadherin=24.02,FVimentin=57.65,FSnail=87.24,FE-cadherin=135.8,P<0.05).qRT-PCR results showed that with the in-crease of CBG concentration,the mRNA expression levels of PI3K,AKT,N-cadherin,Vimentin and Snail de-creased,while the mRNA expression level of E-cadherin gradually increased(FPI3K=101.1,FAKT=558.3,FN-cadherin=86.97,FVimentin=105.9,FSnail=85.71,FE-cadherin=80.96,P<0.01).Conclusion CBG can reverse cisplatin resistance of ovarian cancer A2780/DDP cell line,and its mechanism may be related to the regulation of PI3K/AKT signaling pathway and inhibition of EMT by CBG.
cinobufaginovarian cancercisplatin resistancereversal of drug resistancePI3K/AKTEMTmecha-nism of action
万方数据
维普
