基于网络药理学及实验验证探讨灵芝抗胃癌的分子机制

Molecular mechanism of Ganoderma against gastric cancer based on network pharmacology and experimental test

基于网络药理学、分子对接技术及细胞实验预测灵芝抗胃癌的分子机制。利用中药系统药理学分析平台(traditional Chinese medicine systems pharmacology database and analysis platform,TCMSP)收集灵芝的活性成分和作用靶点。通过GeneCards和OMIM数据库获取胃癌相关的靶点。筛选获得两者的共同靶标后,利用STRING数据库进行构建蛋白互作网络,并且利用Bioconductor平台和R语言进行基因本体(Gene Ontology,GO)生物进程及京都基因与基因组百科全书(Kyoto Encyclopedia of Genes and Genomes,KEGG)通路分析。同时应用Cytoscape软件构建"草药-疾病-成分-靶点"网络和"草药-疾病-成分-靶点-通路"网络。另外选取灵芝的关键活性成分β-谷甾醇与蛋白互作网络前15个靶标进行分子对接。最后利用细胞实验进一步验证上述结果。该实验分别获取灵芝活性成分14个,灵芝相关靶点28个,并且收集灵芝与胃癌的共同靶点25个,包括caspase-3(CASP3)、caspase-8(CASP8)、caspase-9(CASP9)和B-cell lymphoma-2(BCL2)等。KEGG通路分析筛选了72条相关信号通路,显示细胞凋亡和p53信号通路可能在灵芝抗胃癌的过程中起关键作用。分子对接结果表明β-谷甾醇与蛋白互作网络前15个靶标对接良好。细胞实验表明β-谷甾醇通过调控细胞凋亡和细胞周期,有效抑制人胃癌细胞AGS增殖,这可能与p53信号通路有部分关系。以上结果表明,灵芝抗胃癌具有多成分、多靶点、多通路协同作用的特点,为后续深入研究灵芝抗胃癌的复杂机制提供了理论依据。

This study aims to explore the molecular mechanism of Ganoderma against gastric cancer based on network pharmacology,molecular docking,and cell experiment.The active components and targets of Ganoderma were retrieved from Traditional Chinese Medicine Systems Pharmacology Database and Analysis Platform(TCMSP),and gastric cancer-related targets from GeneCards and Online Mendelian Inheritance in Man(OMIM).The protein-protein interaction(PPI)network of the common targets was constructed with STRING,followed by Gene Ontology(GO)term enrichment and Kyoto Encyclopedia of Genes and Genomes(KEGG)pathway enrichment analysis of the common genes based on Bioconductor and R language.The medicinal-disease-component-target network and medicinal-disease-component-target-pathway network were established by Cytoscape.Molecular docking was performed betweenβ-sitosterol(the key component in Ganoderma)and the top 15 targets in the PPI network.Cell experiment was performed to verify the findings.A total of 14 active components and 28 targets of Ganoderma were retrieved,and the medicinal and the disease shared 25 targets,including caspase-3(CASP3),caspase-8(CASP8),caspase-9(CASP9),and B-cell lymphoma-2(BCL2).The common targets involved 72 signaling pathways and apoptosis and p53 signaling pathway may play a crucial role in the effect of Ganoderma against gastric cancer.β-sitosterol had strong binding activity to the top 15 targets in the PPI network.The in vitro cell experiment demonstrated thatβ-sitosterol inhibited gastric cancer AGS cell proliferation by inducing cell apoptosis and cell cycle arrest in the S phase,which might be related to the regulation of the p53 pathway.This study shows the multi-component,multi-target,and multi-pathway characteristics of Ganoderma against gastric cancer,which lays a scientific basis for further research on the molecular mechanism.