Mechanism of cinobufagin regulating PI3K/AKT signaling pathway to reverse cisplatin resistance in ovarian cancer A2780/DDP cells / 安徽医科大学学报

ABSTRACT

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) . @*Results@#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 ( t P-PI3K/PI3K = 8 . 115 , t P-AKT/AKT = 17. 62 , t N-cadherin = 6. 126 , t Vimentin = 4. 001 , t Snail = 17. 333 , t E-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 , F Vimentin = 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 , F Vimentin = 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.