基于网络药理学和实验验证探究左归丸抑制头颈部鳞状细胞癌的作用机制  

Mechanism of Zuoguiwan in inhibiting head and neck squamous cell carcinoma based on network pharmacology and experimental verification

作　　者：周仕敏 谭雅琴[2] 张丽[2] 邓志鹏 ZHOU Shimin;TAN Yaqin;ZHANG Li;DENG Zhipeng(Department of Stomatology,Dongfeng Maojian Hospital,Shiyan,Hubei 442000,China;Department of Stomatology,Taihe Hospital,Hubei Medical College,Shiyan,Hubei 442000,China)

机构地区：[1]国药东风茅箭医院口腔科,湖北十堰442000 [2]湖北医药学院附属太和医院口腔科,湖北十堰442000

出　　处：《安徽医药》2024年第7期1312-1317,I0001,I0002,共8页Anhui Medical and Pharmaceutical Journal

基　　金：湖北省教育厅科学技术研究项目(Q20222114)。

摘　　要：目的基于网络药理学和体外实验探究左归丸作用于头颈部鳞状细胞癌(HNSCC)的靶点并分析潜在机制。方法自2022年11月至2023年3月分别通过TCMSP、BATMAN-TCM、GeneCards、CTD、OMIM数据库筛选左归丸的活性成分并筛选左归丸作用于HNSCC的潜在靶点;使用String数据库构建潜在靶点互作网络,并进行多步拓扑分析筛选出核心靶点;使用DAVID和Metascape数据库对潜在靶点进行富集分析;最后运用Auto Dock Vina进行核心成分与靶点的分子对接并通过细胞计数试剂盒(CCK-8)、蛋白质印迹法验证左归丸对HNSCC的治疗作用及机制。结果左归丸的8种活性成分具有487个药物靶点,其中440个为HNSCC相关基因;两步拓扑结构分析得到表皮生长因子(EGFR)、细胞周期蛋白D1(CCND1)、热休克蛋白90α家族A类成员1(HSP90AA1)、核因子kappa B激酶亚基β抑制剂(IKBKB)和螺旋环螺旋结构域扩散激酶(CHUK)5个核心靶点;富集结果提示左归丸通过影响多种生物学功能和信号通路在HNSCC中发挥作用;分子对接结果显示,左归丸与核心靶点之间结合稳定。体外实验显示,与对照组相比,左归丸处理组显著抑制了WSU-HN6和CAL-27细胞增殖能力(P<0.001)并且可以降低CCND1,IKBKB和CHUK(P<0.05)的蛋白表达水平。结论左归丸能够靶向多条促癌信号通路从而对HNSCC起到抑制作用。Objective To identify the targets of Zuoguiwan(ZGW)on head and neck squamous cell carcinoma(HNSCC)and to explore the potential mechanism based on network pharmacology and in vitro experiments.Methods From November 2022 to March 2023,TCMSP,BATMAN-TCM,GeneCards,CTD and OMIM were employed to analyze the active components of ZGW and to screen the potential targets of ZGW on HNSCC.String database was used to construct an interaction network for potential targets,and the core targets were obtained via multiple step topology analysis.The DAVID and Metascape databases were used for enrichment analysis of the potential targets.Finally,molecular docking between core components and targets was performed by Auto Dock Vina,and CCK-8 and Western blot were used to verify the therapeutic effect and mechanism of ZGW on HNSCC.Results The 8 active components with 487 drug targets were screened out,440 of which were HNSCC-related genes.Two-step topological data analysis results showed that epidermal growth factor receptor(EGFR),cyclin D1(CCND1),heat shock protein 90 alpha family class A member 1(HSP90AA1),inhibitor of nuclear factor kappa B kinase subunit beta(IKBKB)and component of inhibitor of nuclear factor kappa B kinase complex(CHUK)were the core target genes.The enrichment analysis results suggested that ZGW played a role in HNSCC by regulating various biological functions and signaling pathways.Molecular docking results showed that ZGW docked well with core targets.In vitro experiment results showed that compared with the control group, ZGW treatment groups significantly inhibited the proliferation ability of WSU-HN6 and CAL-27 cells (P<0.001) and reduced the protein expression levels of CCND1, IKBKB and CHUK (P<0.05).Conclu sion ZGW can target multiple cancer-promoting signaling pathways to inhibit the progression of HNSCC.

关 键 词：左归丸 头颈鳞癌 网络药理学 分子对接 作用机制 

分 类 号：R285[医药卫生—中药学]