Prunella Vulgaris Extract Induces Apoptosis In Human Breast Cancer MDA-MB-231 Cells And Its Mechanis

Purpose:By studying the growth effect of Ursolic acid(UA)in Triple-negative breast cancer(TNBC)MDA-MB-231 cells in Triple-negative breast cancer(TNBC),and investigating its effects on MDA-MB-231 cell proliferation,apoptosis and cell cycle through in vitro experiments,and analyzing the mechanism of action of ursolic acid inducing apoptosis,it laid a theoretical basis for drug development.Material and method:In vitro culture of three-negative breast cancer MDA-MB-231 cells in the logarithmic growth stage,after the action of ursolic acid in different concentration gradients(0,10,20,40,60,80μmol/L)for 24 h,48h,72 h,the effect of Ursolic acid on MDA-MB-231 cell activity was determined by cell Counting Kit-8 method(and the effective drug concentration was determined;the effective concentration(0,20,40,60,80μmol/L) ursolic acid acted on MDA-MB-231 cells for 48 h,and the expression levels of apoptotic proteins Bax,Bcl-2 and Caspase-3 were detected by flow cytometry after treatment with effective concentrations of ursolic acid and the effect on the cell cycle;after the effective concentration of ursolic acid was determined by protein imprinting(Western blot)on MDA-MB-231 cells for 48 h,the expression levels of apoptotic-related proteins Bax,Bcl-2 and Caspase-3 were determined.Results:1.The results of the CCK-8 method show that ursolic acid can significantly inhibit the growth of MDA-MB-231 cells in triple-negative breast cancer.Except for the treatment of MDA-MB-231 cells at a concentration of 10 μmol/L at a concentration of 10 μmol/L,the cell survival rate was not significantly reduced compared with the control group(P>0.05),and the MDA-MB-231 cells treated with 0,20,40,60,80 μmol/L ursolic acid gradually decreased with the increase of the drug concentration at the same time of action,and the cell survival rate gradually decreased with the prolongation of the drug action time at the same drug concentration.The difference was statistically significant(P<0.05).When ursolic acid acted on cells at 24 h,48 h,and 72 h,the maximum inhibitory concentrations ic50 of half of the inhibition cell activities were: 61.64 μmol/L,38.83 μmol/L,and 23.15 μmol/L,respectively.2.According to the cell cycle detection results,after 48 h of ursolic acid treatment of 0,20,40,60 and 80 μmol/L,with the increase of ursolic acid concentration,the proportion of MDA-MB-231 cells in the G0/G1 stage gradually increased,and the proportion of MDA-MB-231 cells in the S phase and G2/M phase was gradually decreased,and the difference was statistically significant compared with the control group(P<0.05).3.According to the results of apoptosis detection,after 48 h of treatment with ursolic acid of0,20,40,60 and 80 μmol/L,the apoptosis rate of MDA-MB-231 cells gradually increased with the increase of drug concentration,and the difference was statistically significant(P<0.05).4.According to the test results of Western Blot,after 48 h treatment with ursolic acid of 0,20,40,60 and 80 μmol/L,the expression levels of the pro-apoptotic proteins Bax and Caspase-3 in MDA-MB-231 cells increased,and the expression levels of anti-apoptotic protein Bcl-2 decreased,and the comparative difference between the groups was statistically significant(P<0.05).Conclusion:Ursolic acid has a significant inhibitory effect on the growth of MDA-MB-231 cells in triple-negative breast cancer,and is time-and dose-dependent,which can block the cell cycle in the G0/G1 stage and induce apoptosis,and its pro-apoptotic mechanism may be related to the upregulation of bax and Caspase-3 protein expression,downregulation of the expression of Bcl-2 protein,and activation of mitochondrial-mediated endogenous apoptosis pathways.