温阳解毒方治疗结直肠癌的核心靶基因筛选及其机制分析

Core target gene screening and mechanism analysis of yang-warming and toxin-resolving formula in treatment of colorectal cancer

目的 利用网络药理学及分子对接技术筛选温阳解毒方治疗结直肠癌的核心靶基因,并探讨其可能的作用机制。方法 通过TCMSP、Uniprot数据库检索温阳解毒方6种中药的有效成分及相关靶基因,经Genecards数据库筛选得到结直肠癌相关疾病靶基因,二者取交集后得到温阳解毒方治疗结直肠癌的潜在靶基因。基于STRING数据库、Cytoscape软件绘制蛋白质互相作用网络(PPI)图并进行拓扑学分析,得到温阳解毒方治疗结直肠癌的核心靶基因。对核心靶基因进行基因本体(GO)功能、KEGG通路富集分析,采用分子对接技术对6种中药重复的关键活性成分和核心靶基因进行结构验证。结果 温阳解毒方6种中药的相关靶基因269个,与1 349个结直肠癌相关疾病靶基因取交集后,得到温阳解毒方治疗结直肠癌的潜在靶基因131个。PPI图的拓扑学分析结果显示,温阳解毒方治疗结直肠癌的核心靶基因有6个,分别是丝氨酸/苏氨酸蛋白激酶1(Akt1)、p53、表皮生长因子受体(EGFR)、JUN、雌激素受体1(ESR1)、肿瘤坏死因子(TNF)。GO功能及KEEG富集分析结果显示,核心靶基因主要作用于综合肿瘤信号通路、细胞衰老信号通路、p53信号通路、TNF信号通路、PI3K/Akt信号通路等。分子对接结果显示,Akt1、TNF、EGFR与温阳解毒方6种中药的11种关键活性成分均有较低的结合能,其中TNF与甲基黄连碱、EGFR与β-谷甾醇是结合能最低的两组,结合能分别为-6.78、-6.56。结论 温阳解毒方治疗结直肠癌的核心靶基因有Akt1、p53、JUN、EGFR、ESR1、TNF,其机制可能与调控p53、TNF、PI3K/Akt等信号通路有关。

Objective To identify the core target gene for yang-warming and toxin-resolving formula in treating colorectal cancer by network pharmacology and molecular docking,and to explore the mechanism of action.Methods The effective ingredients and related targets of yang-warming and toxin-resolving formula were screened out through TCMSP and Uniprot database.Colorectal cancer-related targets were obtained through the Genecards database.The intersection of these two datasets yielded potential targets for yang-warming and toxin-resolving formula in treating colorectal cancer.Based on STRING database and Cytoscape software,a protein-protein interaction(PPI)network was constructed,followed by topological analysis to identify the core target for yang-warming and toxin-resolving formula in treating colorectal cancer.Then Gene ontology(GO)function and KEGG pathway enrichment analysis were applied to the core target,and molecular docking was employed for structural validation of the key active ingredients shared by the six Chinese medicines and their core targets.Results By intersecting the 269 related targets of the six Chinese herbal medicines in yang-warming and toxin-resolving formula with the 1349 colorectal cancer-related targets,we discovered 131 potential targets for treating colorectal cancer with yang-warming and toxin-resolving formula,and the topological analysis of the PPI network showed that there were 6 core targets, namely serine/threonine protein kinase 1(Akt1), p53, JUN, epidermal growth factor receptor(EGFR), estrogen receptor 1 (ESR1), and tumor necrosis factor (TNF). The results from GO function and KEGGpathway enrichment analysis indicated that these core targets primarily acted on comprehensive tumor signaling pathways,cell senescence signaling pathways, p53 signaling pathways, TNF signaling pathways, and P13K/Akt signaling pathways,etc. Results from molecular docking demonstrated that low binding affinities were exhibited by Akt1, TNF, EGFR as wellas the 11 key active ingredients of the six kinds of traditional Chinese medicines in the formula, among which, Worenine(methyl berberine) and TNF, and β-sitosterol and EGFR showed the lowest binding affinities, with values of -6. 78 and-6. 56, respectively. Conclusion The core target genes for treating colorectal cancer with yang-warming and toxin-resolvingformula include Akt1, p53, JUN, EGFR, ESR1, and TNF, and the mechanism may be related to the regulation ofp53, TNF, P13K/Akt, and other signaling pathways.