Exploration of the Mechanism of EGCG Against MDA-MB-231 cells based on Network Pharmacology
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摘要: 基于网络药理学分析绿茶中的主要多酚物质表没食子儿茶素没食子酸酯 (Epigallocatechin gallate, EGCG)对三阴性乳腺癌细胞MDA-MB-231的潜在作用靶点及其分子机理。使用数据库检索EGCG和MDA-MB-231的潜在靶点，两者的靶点基因相互映射取交集，获得共同作用靶点；借助Cytoscape 3.8.0软件绘制“靶点-通路”网络互作图、String数据库构建靶点蛋白的互作网络关系 (Protein-protein interaction，PPI)，基于Metascape平台对靶点进行基因本体论 (Gene Ontology，GO) 和京都基因与基因组百科全书 (Kyoto Encyclopedia of Genes and Genomes，KEGG) 富集分析；通过分子对接和体外实验验证预测结果。结果表明：通过挖掘，共获得EGCG靶点537个，MDA-MB-231靶点181个，取交集获得88个共同潜在靶点，进一步筛选保留30个核心作用靶点；获得20条核心GO生物进程和17条KEGG信号通路，涉及到癌症信号通路，毒性耐受通路，胰腺癌通路、直肠癌通路，小细胞肺癌通路等；分子对接结果显示，EGCG可以通过非共价键与β-连环蛋白 (β-catenin) 结合；体外实验表明，肝细胞生长因子 (Hepatocyte growth factor，HGF) 能够诱导β-catenin的表达，而EGCG能够抑制HGF诱导的β-catenin的表达上调。EGCG可以通过多靶点、多途径干预MDA-MB-231，其中，已经初步证实EGCG可以影响HGF/β-catenin途径，为进一步机理探究提供理论和实践基础。
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关键词:
- 表没食子儿茶素没食子酸酯 (EGCG) /
- 三阴性乳腺癌 /
- 网络药理学 /
- 分子对接 /
- 作用机理
Abstract:ObjectiveTo screen out the potential targets and molecular mechanisms of Epigallocatechin gallate (EGCG) in the treatment of triple negative breast cancer (MDA-MB-231). MethodDatabases was used to explore the potential targets between EGCG and MDA-MB-231. The “target-pathway” networks of common targets were constructed using Cytoscape 3.8.0 software, while the String database was used to draw and analyze the PPI network. Subsequently, the core genes were submitted to the Metascape database for GO and KEGG enrichment analysis; Finally, the prediction results were verified through the in vitro experiment. ResultsIn our study, a total of 537 EGCG targets and 181 disaster targets were obtained, 30 key targets were retained by further screening from 88 common potential targets. The result of the enrichment analysis showed that the active targets were involved in 20 core GO biological processes and 17 KEGG signaling pathways, including cancer signaling pathways, toxic tolerance pathways, pancreatic cancer pathways, rectal cancer pathways, small cell lung cancer pathways and so on. Molecular docking illuminated that EGCG could interact with β-catenin in a non-covalent manner. The in vitro experiment revealed that HGF could induce the expression of β-catenin, and EGCG could repress the HGF-induced over-expression of β-catenin. ConclusionEGCG inhibits cell viability through multiple targets and multiple pathways, among them, it has been initially confirmed that EGCG can affect the HGF / β-catenin pathway, providing a theoretical and practical basis for further mechanism exploration. -
图 1 EGCG和MDA-MB-231共同潜在靶点筛选
Figure 1. Screening of common and potential targets for EGCG and MDA-MB-231
图 7 Western Blot 检测β-catenin蛋白的表达
Figure 7. The expression of β-catenin identified by Western Blot analysis
表 1 EGCG和MDA-MB-231共同潜在靶点
Table 1. Common and potential targets for EGCG and MDA-MB-231
Gene Protein name MAPK14 Mitogen-activated protein kinase 14 AKT1 RAC-alpha serine/threonine-protein kinase MAPK3 Mitogen-activated protein kinase 3 SP1 Transcription factor Sp1 MAPK8 Mitogen-activated protein kinase 8 SRC Proto-oncogene tyrosine-protein kinase Src IGF1 Insulin-like growth factor I VEGFA Vascular endothelial growth factor A MET Hepatocyte growth factor receptor JUN Transcription factor AP-1 HGF Hepatocyte growth factor SMAD3 Mothers against decapentaplegic homolog 3 FOS Proto-oncogene c-Fos RB1 Retinoblastoma-associated protein ITGB1 Integrin beta-1 PTK2 Focal adhesion kinase 1 RELA Transcription factor p65 PIK3CA Phosphatidylinositol 3-kinase catalytic subunit alpha isoform BCL2L1 Bcl-2-like protein 1 STAT3 Signal transducer and activator of transcription 3 ESR1 Estrogen receptor NFKB1 Nuclear factor NF-kappa-B p105 subunit MAPK1 Mitogen-activated protein kinase 1 CDKN1A Cyclin-dependent kinase inhibitor 1 EGFR Epidermal growth factor receptor EGF Epidermal growth factor CTNNB1 Catenin beta-1 BCL2 Apoptosis regulator Bcl-2 CDH1 Cadherin-1 TP53 Cellular tumor antigen p53 
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