Apoptosis And Mechanism Of Brucein On Triple-negative Breast Cancer Cells

Purpose:In order to explore the inhibitory effect of Chinese herb brucea extract(brusatol)on the growth of triple negative breast cancer MDA-MB-231 cells and the effect of brusatol on the apoptosis of tumor cells and to explore the possible molecular mechanism of inducing the apoptosis of MDA-MB-231 cells.Based on the experimental results obtained,we will explore the drugs that can effectively prevent and treat triple-negative breast cancer in clinic.Material and method : Cultured triple negative breast cancer MDA-MB-231 cell,MDA-MB-231 cells were treated with 0,5,10,20,40 and 80?mol/L Bru for 24,48 and72 h,CCK-8 assay kit was used to determine the growth inhibition rate of MDA-MB-231 cells.The morphological changes of MDA-MB-231 cells after drug treatment were recorded by microscope,Flow cytometry was used to detect the apoptotic effect of brusatol on MDA-MB-231 cells of triple negative breast cancer,The expression levels of related apoptotic proteins Bax,Bcl-2,Cytochrome C and Caspase-3 in MDA-MB-231 cells were detected by Western blot.Results:1.CCK-8 results showed that brusatol could significantly inhibit the growth of MDA-MB-231 cells in triple negative breast cancer.The maximum inhibition rate was81.69%.In addition,There was no significant increase in the inhibitory rate of brusatol at40?mol/L and 80?mol/L for 48 h and 72 h with the increase of time or concentration(P>0.05),In addition,comparison among all groups showed that under the same action time,The inhibitory rate of brusatol on MDA-MB-231 cells increased with the increase of the concentration of brusatol,and there was a significant difference between different concentration groups(P<0.05),which was of statistical significance;Under the same drug concentration,the inhibition rate of MDA-MB-231 cells also increased with the prolongation of drug action time,and there was a significant difference between the groups(P<0.05),which was statistically significant;2.The cells were treated with 0,5,10,20 and 40?mol/L BRU for 48 hours.Under the microscope observation,compared with the cell morphology without the addition of the drug,The cell morphology of MDA-MB-231 showed an irregular state such as elongated or dendritic,increased cell space,poor adherent state,and many suspended cells and other apoptotic states.3.The results of flow cytometry indicated that brusatol could induce apoptosis in MDA-MB-231 cells.After treated with 0,5,10,20,40 ?mol/L BRU for 48 h,The higher the Bru concentration,the higher the apoptosis rate,and there was a significant difference among groups,with statistical significance(P<0.05).4.The cells were treated with 0,5,10,20 and 40?mol/L Bru for 48 hours,Western blot analysis showed that brusatol could induce apoptosis of MDA-MB-231 cells,which may be induced by the activation of endogenous mitochondrial proteins,Among them,t The expression levels of Bax,Cytochrome C and Caspase 3,which promote apoptosis,were significantly increased,while the expression levels of Bcl-2,which inhibit apoptosis,were decreased,And with the increase of drug concentration,the corresponding trend of change also increased,And there were significant differences among the concentration groups,with statistical significance(P<0.05).These results indicated that MDA-MB-231 cells with triple negative breast cancer could undergo apoptosis after treatment with brusatol,which might be caused by the activation of mitochondrial apoptotic pathway related proteins.Conclusion:According to the results,brusatol can significantly inhibit the growth of MDA-MB-231 cells in triple negative breast cancer,and promote the changes of cell morphology,and then induce cell apoptosis.It may be that the protein expressions of cytochrome C,Bax and Caspase 3 are increased and the expression of Bcl-2 is decreased by activating the related proteins of mitochondrial apoptosis pathway,thus accelerating the apoptosis of cells.