Study On The Inhibitory Effect Of 27-O-p-(E)-coumaroyl Ursolic Acid In The Leaves Of Ilex Latifolia On Breast Cancer Cells And Its Mechanism

Objective: To investigate the effect of triterpenoid 27-O-p-(E)-coumaroyl ursolic acid(27-CAUA)in the leaves of Ilex latifolia on the growth of breast cancer cells and its intervention in the pathway of programmed cell death.Methods: The MTT method was used to compare the inhibitory effects of 27-CAUA on the proliferation of different breast cancer cell lines and normal breast cells.Observe the effect of 27-CAUA on the morphology of different breast cancer cell lines and normal breast cells;use immunofluorescence to detect the expression and localization of 27-CAUA on the intracellular proteins EGFR,Her2,LC3A/B,BRUCE and Cleaved-Caspase-3 impact.Flow cytometry was used to detect the expression of27-CAUA on the intracellular protein Her2 and LC3A/B and the effect of 27-CAUA on the apoptosis signal pathway of breast cancer cells.The m RFP-GFP-LC3 tandem fluorescent protein adenovirus was used to infect breast cancer cells to observe the effect of 27-CAUA on the autophagy signaling pathway of breast cancer cells.Result:1.27-CAUA has a certain dose-dependent inhibitory effect on the growth of MDA-MB-468,MDA-MB-435 S and HCC1806 cells.On normal human breast cells MCF-10 A,it shows a trend of first promoting and then inhibiting as the dose increases..2.EGFR protein test results showed that human breast cells MCF-10 A EGFR expressed,MAD-MB-468 cells EGFR overexpressed,MDA-MB-435 S cells EGFR low expression,HCC1806 cells did not express EGFR.3.27-CAUA interferes with the migration of Her2 in MDA-MB-468 cells and HCC1806 cells,resulting in changes in the pathway of programmed cell death.4.27-CAUA can induce an increase in the expression of BRUCE and LC3A/B in HCC1806 cells,and the cells transfected with adenovirus have obvious autophagosome-lysosome fusion,indicating that 27-CAUA can induce autophagic lysis of HCC1806 cells Enzyme body formation.27-CAUA cannot induce autophagolysosome formation in MDA-MB-468 cells.5.27-CAUA can induce the increase of Cleaved Caspase-3 expression in MDA-MB-468 cells.Annexin-V/PI staining flow cytometry test showed that27-CAUA can induce apoptosis of MDA-MB-468 cells,but 27-CAUA has no effect on promoting apoptosis of HCC1806 cells.Conclusion:1.The triterpenoid 27-CAUA from leaves of Ilex latifolia induced programmed death of breast cancer cells MDA-MB-468,HCC1806 and MDA-MB-435 S in a dose-dependent manner.2.27-CAUA promotes the metastasis of Her2 in breast cancer cells overexpressing EGFR,and also promotes the metastasis of Her2 in breast cancer cells that do not express EGFR,but the direction of movement is affected by EGFR.3.27-CAUA induced the displacement of Her2,which led to the programmed death of different breast cancer cells,and led to changes in the pathways of programmed death of breast cancer cells.