基于网络药理学及分子对接与实验验证苗药抑癌汤经WNT/β-catenin信号通路抑制小细胞肺癌增殖的作用机制  

Based on Network Pharmacology and Molecular Docking and Experimental Verification of theMechanism of Miao-Yi-Ai-Tang Inhibiting the Proliferation of Small Cell Lung Cancer throughWNT/β-Catenin Signaling Pathway

作　　者：陈珊 李波 葛正行 谭韬 张军 余梅 龚香群 CHEN Shan;LI Bo;GE Zhengxing;TAN Tao;ZHANG Jun;YU Mei;GONG Xiangqun(The Second Clinical Medical College of Guizhou University of Traditional Chinese Medicine,Guizhou 550002,China;The Second Affiliated Hospital of Guizhou University of Traditional Chinese Medicine,Guizhou 550001,China)

机构地区：[1]贵州中医药大学第二临床医学院,贵州550002 [2]贵州中医药大学第二附属医院,贵州550001

出　　处：《世界科学技术-中医药现代化》2024年第7期1847-1861,共15页Modernization of Traditional Chinese Medicine and Materia Medica-World Science and Technology

基　　金：贵州省科学技术厅局级项目(黔科合基础[2016]2013):苗药抑癌方基于WNT/β-atenin信号通路调控小细胞肺癌细胞株NCI-H446增殖抑制机制研究,负责人:李波;贵州中医药大学研究生教育创新计划项目(YCXKYS2023012):基于网络药理学及实验验证苗药抑癌汤联合顺铂经PI3K/Akt/mTOR通路抑制小细胞肺癌增殖的研究,负责人:李波。

摘　　要：目的运用网络药理学及分子对接技术挖掘并预测苗药抑癌汤(Miao-Yi-Ai-Tang,MYAT)治疗小细胞肺癌(Small cell lung cancer,SCLC)的作用靶点及相关信号通路,并通过细胞及动物实验验证其作用机制,为基础实验及临床应用提供理论依据。方法从TCMSP数据库获取MYAT有效成分,并结合PubMed、Swiss Target Prediction及Uniprot数据库获取潜在靶点;通过DrugBank、Genecards、OMIM、TTD数据库收集SCLC相关基因,通过Venny 2.1平台获取MYAT和SCLC的交集基因后导入STRING数据库,构建蛋白质相互作用网络(Protein-protein interaction,PPI),运用Cytoscape 3.9.1软件进行可视化分析,利用Metascape数据库进行GO富集分析及KEGG通路分析,预测MYAT治疗SCLC的直接作用靶点及信号通路;使用AutoDock Tools 1.5.7软件进行分子对接验证两者之间的紧密联系。通过细胞学实验验证,培养细胞加入MYAT处理并使用qPCR检测β-catenin、AXIN、c-myc的表达,Western blot检测细胞中β-catenin的表达;动物实验建立肺癌NCI-H446皮下移植瘤模型,观察MYAT对肿瘤生长的影响。结果共获得MYAT有效成分65个,SCLC基因1368个,MYAT与SCLC交集基因260个,富集分析显示与癌症通路、PD-L1/PD-1通路、NF-κB通路、Wnt等信号通路有关。分子对接验证结果显示,其有效活性成分与核心靶点蛋白的结合能均<0 kJ·mol-1,说明蛋白可与活性成分自发结合且稳定性较好。细胞实验显示MYAT组β-catenin、c-myc及AXIN mRNA表达量均显著下调(P<0.05)。动物实验显示:MYAT能明显抑制肿瘤在体内的生长。结论苗药抑癌汤可通过Wnt/β-catenin信号通路抑制小细胞肺癌增殖。Objective To use network pharmacology to mine and predict the targets and related signaling pathways of Miaoyao Yiai Tang(Miao-Yi-Ai-Tang,MYAT)in the treatment of small cell lung cancer(SCLC).And animal experiments to verify its mechanism of action,to provide a theoretical basis for basic experiments and clinical applications.Methods The active ingredients of MYAT were obtained from the TCMSP database,combined with PubMed data,Swiss Target Prediction database and Uniprot database to obtain potential targets;SCLC-related genes were collected through the DrugBank database,Genecards database,OMIM database and TTD database,and the Venny 2.1 platform After obtaining the intersection genes of MYAT and SCLC,import them into the STRING database,construct a protein-protein interaction(PPI)network,use Cytoscape 3.9.1 software for visual analysis,and use Metascape database for GO enrichment analysis and KEGG pathway analysis,to predict the direct action target and signaling pathway of MYAT in the treatment of SCLC.Using AutoDock Tools 1.5.7 software for molecular docking to verify the close relationship between the two.For cytological experiment verification,the cultured cells were treated with MYAT and the expression ofβ-catenin,AXIN,c-myc was detected by qPCR,and the expression ofβ-catenin in the cells was detected by Western blot;animal experiments were established to establish a subcutaneous xenograft tumor model of lung cancer NCI-H446,to observe the effect of MYAT on tumor growth.Results A total of 65 effective components of MYAT,1368 SCLC genes,and 260 MYAT-SCLC intersection genes were obtained.Enrichment analysis showed that they were related to cancer pathways,PD-L1/PD-1 pathways,NF-κB pathways,Wnt and other signaling pathways.The results of molecular docking validation showed that the binding energies of active components and core target proteins were all<0 kJ·mol-1,which indicated that the protein could spontaneously bind to active components and be stable.Cell experiments showed that the expression levels of

关 键 词：苗药抑癌汤 网络药理学 分子对接 WNT/Β-CATENIN信号通路 小细胞肺癌(SCLC) NCIH446 细胞 

分 类 号：R285.5[医药卫生—中药学]