Effects Of Phloretin Independ Or Combine With 3-MA On Proliferation And Apoptosis Of CAOV3 Cells

ObjectiveTo observe the effects of different concentrations of Phloretin on the proliferation and apoptosis of CAOV3 cells and to explore its mechanism,and to observe the effect of Phloretin combined with autophagy inhibitor 3-MA on the proliferation and apoptosis of CAOV3 cells.It provides reliable experimental data for the use of Phloretin combined with autophagy inhibitor in the in vitro cell experiment of ovarian cancer.MethodsMTT assay was used to detect the inhibitory effect of different concentrations of Phloretin on the proliferation of CAOV3 cells,and flow cytometry was used to detect the changes of apoptosis rate of CAOV3 cells treated with different concentrations of Phloretin.Western blot was used to detect the expression of apoptotic proteins BAX,p53 and p38 signal transduction pathway related proteins in CAOV3 cells treated with different concentrations of Phloretin.Western blot was used to detect the expression of autophagy labeled protein LC3 B in CAOV3 cells treated with Phloretin alone and combined with 3-MA,and MTT assay was used to detect the inhibitory effect of Phloretin alone and combined with 3-MA on the proliferation of CAOV3 cells.Flow cytometry was used to detect the apoptosis rate of CAOV3 cells treated with Phloretin alone and combined with 3-MA.Results1.Phloretin could inhibit the growth of ovarian cancer CAOV3 cells in a dose-time dependent manner,which was significantly higher than that of the control group(P < 0.01).2.The apoptosis rate of ovarian cancer CAOV3 cells increased with the increase of the concentration of Phloretin and the expression of apoptotic protein BAX and p53 also increased(P < 0.05).3.The results showed that the expression of p-p38 protein increased gradually with the increase of dose,but the expression concentration of p38 protein did not change significantly in ovarian cancer CAOV3 cells treated with different concentrations of Phloretin.There was no significant difference between the two groups(P > 0.05).4.The autophagy system was activated during the apoptosis of ovarian cancer CAOV3 cells induced by Phloretin.the results showed that the expression of autophagy labeling protein LC3B-?decreased and LC3B-? increased(P < 0.05).The autophagy was inhibited after administration of 3-MA,an autophagy inhibitor.5.The combination of Phloretin and inhibitor 3-MA could significantly inhibit the growth of CAOV3 cells and increase the apoptosis rate of CAOV3 cells compared with the control group(P < 0.05).The growth of CAOV3 cells was significantly inhibited and the apoptosis rate of CAOV3 cells was significantly increased compared with the control group(P < 0.05).ConclusionsPhloretin can inhibit the proliferation of CAOV3 cells in a dose-time dependent manner and induce apoptosis of CAOV3 cells in a dose-dependent manner,and its apoptosis mechanism may be regulated by p38 signal transduction pathways.The autophagy system was activated during the treatment of CAOV3 cells with Phloretin.Combined with 3-MA inhibition of autophagy could increase the inhibition rate and apoptosis rate of CAOV3 cells induced by Phloretin.To provide reliable experimental data for the use of Phloretin combined with autophagy inhibitor in tumor cell experiment in vitro.