背景:Hispolon,从药用黄色真菌桑树中分离出的酚类化合物,表现出强大的抗三阴性乳腺癌(TNBC)作用。然而,Hispolon的抗肿瘤机制尚未得到充分探索。
目的:在本研究中,我们基于生物信息学和体外实验,系统地研究了Hispolon抗TNBC的机制。
方法:首先从SwissTarget数据库中收集Hispolon相关目标。使用基因表达综合(GEO)数据集在TNBC和正常乳腺组织之间筛选差异表达基因(DEG)。通过绘制维恩图分析了Hispolon和DEG之间的重叠目标。构建蛋白质-蛋白质相互作用(PPI)网络以分析这些靶标之间的相互作用。重点是通过Cytoscape3.7.2软件的Cytohubba和MCODE插件挖掘Hispolon的抗TNBC效应的核心目标。我们对这些核心目标进行了生存分析,以筛选最匹配的目标,包括EGFR,KIT,和PLAU。这与我们通过分子对接对Hispolon的验证密切相关。此外,使用R软件(ClusterProfiler软件包)进行基因本体(GO)分析和KEGG通路分析。最后,进行体外实验以评估预测的靶基因的准确性.
结果:ADME结果表明Hispolon具有开发成药物的巨大潜力。通过匹配Hispolon的107个靶标与TNBCDEG的2,013个靶标来筛选20个重叠靶标。最初确定了Hispolon对抗TNBC的七个核心目标,包括EGFR,IGFBP3,MMP9,MMP2,MMP1,PLAU,和KIT。GO富集分析表明,Hispolon作用于TNBC的生物学过程主要涉及免疫应答中的淋巴细胞活化和磷脂酰肌醇介导的信号转导。此外,松弛素信号通路,雌激素信号通路,蛋白聚糖在癌症中,和其他可能是Hispolon对抗TNBC的关键途径。此外,Hispo-lon以浓度依赖的方式抑制MDA-MB-231细胞的增殖,并调节核心靶标EGFR的RNA和蛋白质表达,部队,和KIT用于治疗TNBC。
结论:在这项研究中,通过网络药理学和体外实验,探讨了Hispolon抗TNBC的多基因药理作用机制,为中药单体抗TNBC的作用机制提供了新的见解。
BACKGROUND: Hispolon, a phenolic compound isolated from the medicinal yellow fungal mulberry, exhibits a strong anti-triple-negative breast cancer (TNBC) effect. However, the antitumor mechanisms of Hispolon have not been fully explored.
OBJECTIVE: In this study, we systematically investigated the mechanism of Hispolon against TNBC based on bioinformatics and in vitro experiments.
METHODS: The Hispolon-related targets were first collected from the SwissTarget database. Differential Expression Genes (DEG) were screened between TNBC and normal breast tissue using the Gene Expression Comprehensive (GEO) dataset. The overlapping targets between Hispolon and DEG were analyzed by plotting Venn maps. Protein-protein interaction (PPI) network was constructed to analyze the interactions among these targets. The focus was on mining the core targets of anti-TNBC effects of Hispolon via the Cytohubba and MCODE plugin of Cytoscape 3.7.2 software. We performed survival analysis on these core targets to screen the best-matched targets, including EGFR, KIT, and PLAU. This correlated strongly with our validation of Hispolon by molecular docking. In addition, Gene Ontology (GO) anal-ysis and KEGG pathway analysis were performed using R software (ClusterProfiler package). Finally, in vitro experiments were performed to assess the accuracy of predicted target genes.
RESULTS: The ADME results suggested that Hispolon has great potential to develop into a drug. Twenty overlapping targets were screened by matching the 107 targets of Hispolon to the 2,013 targets of TNBC DEG. Seven core targets of Hispolon against TNBC were initially identified, including EGFR, IGFBP3, MMP9, MMP2, MMP1, PLAU, and KIT. GO enrich-ment analysis demonstrated that the biological process of Hispolon acting on TNBC mainly involves lymphocyte activation in immune response and phosphatidylinositol-mediated signal-ing. Additionally, the relaxin signaling pathway, estrogen signaling pathway, proteoglycans in cancer, and others might be the key pathways of Hispolon against TNBC. Furthermore, Hispo-lon inhibited the proliferation of MDA-MB-231 cells in a concentration-dependent manner and regulated the RNA and protein expression of the core targets EGFR, PLAU, and KIT for the treatment of TNBC.
CONCLUSIONS: In this study, the polygenic pharmacological mechanism of action of Hispolon against TNBC was explored through network pharmacology and in vitro experiments, provid-ing a new insight into the mechanism of TCM monomer against TNBC.