Exploring the therapeutic potential and in vitro validation of Baicalin for the treatment of triple-negative breast cancer

Yuan Ma¹Ying Pan²乾程 赵³Chongheng Zhang¹Haitao He²Lihua Pan²Jianling Jia⁴Aiping Shi⁴Yiming Yang^3*Wenfeng Zhang¹

• ¹Changchun University of Traditional Chinese Medicine， School of Basic Medicine, Changchun, China
• ²College of Basic Medical Sciences, Jilin University, Changchun, Jilin Province, China
• ³Department of Cell Biology, College of Basic Medical Sciences, Jilin University, Changchun, China
• ⁴Department of Breast surgery, General Surgery Center, the First Hospital of Jilin University, Changchun, China

Objective: To explore the mechanism of action of baicalin (BA) in the treatment of triple-negative breast cancer (TNBC) based on network pharmacology, molecular docking and molecular dynamics simulations and in vitro validation.Methods: The inhibitory effects of different concentrations of baicalin on the proliferation of MDA-MB-231, 4T1, MCF-7, and MCF-10A cell lines were evaluated by CCK8 assay with clone formation assay. Three compound target prediction platforms, Swiss Target Prediction, SEA and Pharmmapper, were used to predict baicalein-related targets, and mapped with the TNBC related targets retrieved from GeneCards and OMMI databases to obtain the potential targets of baicalein for the treatment of TNBC; the STRING database and the STRING database and Cytoscape software were used to construct the protein interaction network and screen the core targets; GO and KEGG enrichment analyses were performed on the core targets; the binding of baicalin to the key targets of TNBC was verified by molecular docking and molecular dynamics simulation; and the expression of the relevant proteins was verified.Results: Baicalin showed more obvious antiproliferative effects on TNBC cell lines at certain concentrations, and had less effect on the proliferation of normal breast cells. A total of 9 core targets of baicalein in the treatment of TNBC, including AKT1, ESR1, TNF-α, SRC, EGFR, MMP9, JAK2, PPARG, and GSK3B, were identified through the construction of the PPI protein interactions network and the ‘Traditional Chinese Medicine-Component-Target-Disease’ network, and a total of 252 targets related to the intersected targets were identified in the GO analysis. GO analysis enriched a total of 2526 Biological process, 105 Cellular component and 250 Molecular function related to the intersecting targets; KEGG analysis enriched a total of 128 signaling pathways related to the intersecting targets; molecular docking results and molecular dynamics studies found that baicalein was able to interact with MMP9, TNF-α, JAK2, PPARG, GSK3B, and other core targets of baicalein for the treatment of TNBC, MMP9, TNF-α, and JAK2 target proteins, and had significant changes in the expression levels of the target proteins.Conclusion: Baicalin inhibits the protein expression of MMP9, TNF-α and JAK2 and their related signaling pathways in the treatment of TNBC.