Effect Of Oridonin And EZH2 Inhibitor GSK126 On Proliferation And Apoptosis Of Ovarian Cancer SKOV3 Cells

ObjectiveThe purpose of this study is to explore the effect of oridonin combined with EZH2 inhibitor GSK126 to enhance the effects of oridonin on the proliferation and apoptosis of ovarian cancer SKOV3 cells and its possible related mechanisms.Methods1.Treat ovarian cancer cells with different concentrations of oridonin and GSK126 for 24 h,48h,72 h,and use MTT experiment to detect the effects of the two drugs on cell viability.According to the results of MTT,the 50% inhibitory concentration of oridonin on SKOV3 cells was screened out,and then the cells were treated with oridonin at different concentrations of GSK126 in combination with a determined concentration for 48 h.After detecting its cell activity and determining the medicinal concentrations of the two drugs,further experiments were carried out in accordance with the control group(no drug group),GSK126 group,oridonin group and combination group.2.Detect the effects of four different treatment groups on cell proliferation through the clone formation experiment.3.Observe the effects of four different treatment groups on the apoptosis of ovarian cancer cells by Hoechst33342 staining.4.Detect the influence of four different treatment groups on cell apoptosis rate by flow cytometry.5.Detect the expression of Bcl-2,Bax and Cleaved-caspase3 related to ovarian cancer cell apoptosis by the combined drug by western blot.6.Detect the expression of AKT,p-AKT,ERK,and p-ERK related pathway proteins in ovarian cancer cells by western blot.Results1.Both oridonin and GSK126 have effective inhibitory effects on SKOV3 cell proliferation and are time-and dose-dependent.The IC50 value of oridonin combined with GSK126 for 48 h is determined to be oridonin 10(μmol /L)GSK126is 20(μmol/L).2.Compared with the control group and the single-drug group,oridonin combined with GSK126 can effectively inhibit the colony formation of SKOV3 cells(P<0.05).3.Hoechst33342 staining showed that compared with the control group and the single drug group,the number of apoptotic cells in the drug group increased the most.4.Through flow cytometry experiments,it was found that the apoptotic cell rate of oridonin combined with GSK126 was higher than that of the control group and single drug foot(P<0.05).5.Western blot results showed that the expression of related apoptosis protein Bcl-2 in SKOV3 cells of the single-drug group was reduced,and the expression of the combined drug group was the lowest.Compared with the control group,the single-drug group had the expression of Bax and Cleaved-caspase3.The amount increased,and the protein expression in the combination group was the highest(P<0.05).6.The results of western blot showed that the relative protein expression levels of p-AKT/AKT and p-ERK/ERK in the PI3K/AKT and MEK/ERK pathways of the combination group were significantly lower than those of the control group(P<0.05).ConclusionsThe combined drug group is more effective than the single drug group in inhibiting the proliferation of ovarian cancer SKOV3 cells and promoting cell apoptosis,enhancing the medicinal efficacy of Rubescensine A on ovarian cancer cells.The mechanism of promoting apoptosis may be related to PI3K/AKT and MEK/ERK pathways.