基于网络药理学、分子对接及实验验证探讨大黄素治疗乳腺癌合并高脂血症的作用机制
Mechanism of Emodin in Treating Breast Cancer Complicated with Hyperlipidemia Based on Network Pharmacology,Molecular Docking,and Experimental Verification
张素方 1李海霞 2刘清清 1王大伟 3刘擎1
作者信息
- 1. 广州中医药大学第二附属医院珠海医院,广东 珠海 519015
- 2. 岳阳新华达制药有限公司,湖南 岳阳 414000
- 3. 广州中医药大学第一附属医院,广东 广州 510405
- 折叠
摘要
目的 基于网络药理学初步预测大黄素治疗乳腺癌合并高脂血症的作用靶点及信号通路,通过分子对接和体外实验验证探讨其可能的作用机制.方法 通过中药系统药理学数据库及分析平台(Traditional Chinese Medicine Systems Pharmacology Database and Analysis Platform,TCMSP)、Swiss Target Prediction数据库查询大黄素的作用靶点;通过GeneCards、OMIM数据库获取乳腺癌、高脂血症的相关靶点;通过在线Veny软件获得大黄素-乳腺癌-高脂血症的共同靶点,并将共同靶点导入STRING数据库构建靶点蛋白质相互作用(protein-protein interaction,PPI)网络,再利用 Cytoscape 3.9.1软件进行可视化处理,筛选核心靶点.通过Metascape在线基因功能注释分析工具进行基因本体(gene ontology,GO)生物学过程和京都基因与基因组百科全书(Kyoto encyclopedia of genes and genomes,KEGG)通路富集分析.选取关键靶点与大黄素通过AutoDock Vina等软件进行分子对接验证.在乳腺癌4T1细胞和肿瘤相关巨噬细胞(tumor-associated macrophage,TAM)中检测大黄素在抑制乳腺癌发展中的作用.以CCK-8法检测大黄素对4T1细胞生长的影响并筛选最佳药物作用浓度,分别采用Real-time PCR及ELISA检测基质金属蛋白酶9(matrix metalloproteinase,MMP9)、白细胞介素-1β(interleukin-1 beta,IL-1β)及转化生长因子β1(transforming growth factor beta 1,TGF-β1)的表达水平.结果 共筛选得到43个大黄素-乳腺癌合并高脂血症共同靶点,核心靶点有肿瘤抗原p53(tumor protein p53,TP53)、肿瘤坏死因子(tumor necrosis factor,TNF)、髓细胞增生原癌基因(myelocytomatosis oncogene,MYC)、表皮生长因子受体(epidermal growth factor receptor,EGFR)、90 kDa热休克蛋白AA1(heat shock protein 90 kDa alpha1,HSP90AA1)、半胱氨酸天冬氨酸蛋白酶3(caspase 3,CASP3)等.涉及癌症通路、化学致癌作用-受体激活、脂质与动脉粥样硬化、膀胱癌、结肠直肠癌、癌症中的蛋白多糖、白细胞介素17(interleukin-17,IL-17)信号通路等.分子对接提示大黄素与核心靶点能较好地结合.实验结果显示,大黄素可显著下调乳腺癌4T1细胞MMP9和TGF-β1的表达.TAM与脂多糖(lipopolysaccharides,LPS)共培养可诱导IL-1β表达,大黄素组可明显下调IL-1β的表达.结论 大黄素治疗乳腺癌合并高脂血症的作用机制可能与抑制肿瘤细胞增殖、转移,诱导肿瘤细胞凋亡及调控肿瘤和高脂血症相关炎性微环境有关.
Abstract
Objective Based on network pharmacology,the targets and signal pathways of emodin in the treatment of breast cancer complicated with hyperlipidemia were preliminically predicted,and the possible mechanisms of emodin were investigated through molecular docking and in vitro experiment.Methods Emodin targets were obtained from the Traditional Chinese Medicine Systems Pharmacology Database and Analysis Platform(TCMSP)and Swiss Target Prediction database.The relevant targets of breast cancer and hyperlipidemia were obtained from GeneCards and OMIM databases,respectively.The common targets of emodin,breast cancer,and hyperlipidemia were obtained using the online software Venny software,and common targets were used to construct a protein-protein interaction(PPI)network in the STRING database.Visual analysis and core target screening were performed using Cytoscape 3.9.1 software.Gene Ontology(GO)biological process and Kyoto Encyclopedia of Genes and Genomes(KEGG)pathway enrichment analysis were conducted using the Metascape online gene function annotation analysis tool.Molecular docking validation was performed using software such as AutoDock Vina for the key targets and emodin.The role of emodin in inhibiting breast cancer development was detected in breast cancer 4T1 cells and tumor-associated macrophage(TAM).The effects of emodin on the growth of 4T1 cells were detected by CCK-8 and the optimal concentration was screened.The expression levels of matrix metalloproteinase 9(MMP9),interleukin-1beta(IL-1β),and transforming growth factor beta 1(TGF-β1)were detected by Real-time PCR and ELISA,respectively.Results A total of 43 common targets related to emodin,breast cancer,and hyperlipidemia were obtained,including core targets such as tumor protein p53(TP53),tumor necrosis factor(TNF),myelocytomatosis oncogene(MYC),epidermal growth factor receptor(EGFR),heat shock protein 90kDa alpha1(HSP90AA1),and caspase 3(CASP3).These targets involved in various pathways such as cancer pathways,chemical carcinogenesis-receptor activation,lipids and atherosclerosis,bladder cancer,colorectal cancer,proteoglycans in cancer,and the interleukin-17(IL-17)signaling pathway.Molecular docking confirmed that emodin exhibited strong binding activity with the core targets.Additionally,experimental results demonstrated that emodin significantly downregulated the expression of MMP9 and TGF-β1 in 4T1 cells.Furthermore,emodin reduced the IL-1β expression induced by co-culture of TAM and lipopolysaccharide(LPS).Conclusion The mechanism of emodin in the treatment of breast cancer complicated with hyperlipidemia may be related to the inhibition of tumor cell proliferation and metastasis,induction of tumor cell apoptosis,and regulation of the inflammatory microenvironment associated with both tumor and hyperlipidemia.
关键词
大黄素/网络药理学/分子对接/乳腺癌/高脂血症Key words
emodin/network pharmacology/molecular docking/breast cancer/hyperlipidemia引用本文复制引用
出版年
2024
国家科技期刊平台
NSTL
万方数据