Effect of pristimerin on adriamycin resistance of osteosarcoma cells by regulating Hippo/YAP signal pathway
Objective To investigate the effect of pristimerin(PM)on adriamycin(ADM)resistance in osteosarcoma cells and its mechanism.Methods The ADM resistant cells MG-63/ADR was established in vitro by using the ADM increasing concentration gradient long-term culture method,CCK-8 was used to detect the effects of different concentrations of ADM on the viability of osteosarcoma cells MG-63 and MG-63/ADR cells to verify the drug resistance of ADM;subsequently,MG-63/ADR cells were randomly divided into the MG-63/ADR group(normal culture),low-dose PM group(adding 0.5 μmol/L PM),middle-dose PM group(adding 1.0 μmol/L PM)and high-dose PM group(adding 2.0 µmol/L PM).The cell viability of each group under different concentrations of ADM treatment was detected by CCK-8;the sensitivity of cells to ADM in each group was detected by plate clone formation assay;the cell morphology of each group was observed under microscope;the cell apoptosis level was detected by flow cytometry;Western blot was used to detect the expression of phosphorylated mammalian STE20-like protein kinase 1(p-MST1),MST1,phosphorylated large tumor suppressor 1(p-LATS1),LATS1,phosphorylated Yes associated protein(p-YAP)and YAP proteins.Results With the intervention of 25,125,625,3 125 and 15 625 ng/mL ADM,the activity of MG-63/ADR cells was significantly higher than that of MG-63 cells(P＜0.05),which indicated that the ADM resistant cell model was successfully established.Compared with the MG-63/ADR group,the number of cells in the low,middle and high-dose PM groups decreased,and the cells gradually shrank,the spacing between cells became larger,and the cell viability,cell clonal formation rate and p-YAP/YAP level decreased(P＜0.05),the apoptosis rate and the levels of p-MST1/MST1 and p-LATS1/LATS1 significantly increased(P＜0.05).Conclusion The inhibitory effect of PM on ADM resistance in osteosarcoma cells may be related to the activation of Hippo pathway and the downregulation of YAP signal.
万方数据
