Study On The Mechanism Of Baicalin On Apoptosis Of Human Triple-negative Breast Cancer Cells MDA-MB-23

Purpose:To analyze and explore the possible targets and molecular mechanism of action of baicalein in the treatment of breast cancer by network pharmacology and molecular docking.In vitro cell experiments demonstrate the effect of scutellarin on the proliferation and apoptosis of triple negative breast cancer cells MDA-MB-231.Material and method:1.The chemical constituents and targets of baicalein were screened using TCMSP and converted into corresponding Gene Symbol,breast cancer disease genes were retrieved from the GeneCards database.The Cytoscape 3.9.0 was used to construct the chemical constituents of scutellaria scutellaria-breast cancer targets network,and the protein interaction(PPI)network of intersection genes was obtained by STRING 11.5 database.The core genes were selected by CytoNCA.GO and KEGG enrichment analysis of intersection genes was performed using DAVID 6.8 database.After that,AutoDockvina was used for molecular docking between the selected compounds and target protein.2.The effects of baicalein on proliferation of human triple-negative breast cancer cell line MDA-MB-231 was measured by CCK-8 assay.After baicalein was treated with 0,6.25,12.5,25,50,100,200,400,800μM for 24 h,the growth inhibition rate of human triple negative breast cancer cells was determined by CCK-8 kit.Effect of baicalein on apoptosis of MDA-MB-231 cells in human triple negative breast cancer was determined by flow cytometry.The expression levels of AKT pathway protein and apoptotic proteins Bcl-2,Bcl-xl and Bax in MDA-MB-231 cells treated with Baicalein were detected by Western blot.Results:1.There are 36 effective components of scutellaria in the treatment of breast cancer through network pharmacology screening,mainly through PI3K-AKT signaling pathway,p53 signaling pathway,MAPK signaling pathway,TNF signaling pathway and IL-17 signaling pathway to play a role in the treatment of breast cancer.Molecular docking showed that baicalein was well docked with TP53,AKT1,CCND1,ESR1 and JUN.2.The 24 h inhibitory rate of baicalin on human triple negative breast cancer MDA-MB-231 cells was 150.21μM and showed a concentration-dependent trend.The higher the concentration,the higher the inhibitory rate.(P < 0.05,with statistical significance)3.Flow cytometry of 100μM and 150μM baicalein on human triple negative breast cancer MDA-MB-231 cells for 24 h showed that the apoptosis rates of 100μM and 150μM baicalein were 26.6% and 30.1%.(P < 0.05,with statistical significance)4.After treating human triple negative breast cancer MDA-MB-231 cells with 0 and 150μM baicalein for 24 h,Western blot assay was used to determine the change of protein content.The apoptosis of human triple negative breast cancer MDA-MB-231 cells was induced by baicalein.It may be caused by the increased expression of apoptotic protein Bax and decreased expression of apoptotic proteins Bcl-2 and Bcl-xl through the inhibition of AKT pathway and downstream related proteins m TOR,SGK1 and 4EBP1.These results indicated that baicalein induced apoptosis of human triple negative breast cancer cells MDA-MB-231 cells by inhibiting the downstream proteins of AKT pathway and regulating apoptosis.(P <0.05,with statistical significance)Conclusion:According to the experimental data,baicalein inhibits the proliferation of human triple negative breast cancer MDA-MB-231 cells and promotes cell apoptosis,which may be caused by inhibiting the expression of apoptosis protein Bax,Bcl-2 and Bcl-xl by inhibiting the related proteins m TOR,SGK1 and 4EBP1 downstream of the AKT pathway.