基于网络药理学和细胞实验探讨贝母素乙抗结肠癌的作用机制  

To Investigate the Mechanism of Peiminine Against Colon Cancer Based on Network Pharmacology and Cell Experiments

作　　者：何勤思 饶军 莫明子 韩晓群[3] 彭紫梅 曾纪权[1] 郑智 易波[1] HE Qinsi;RAO Jun;MO Mingzi;HAN Xiaoqun;PENG Zimei;ZENG Jiquan;ZHENG Zhi;YI Bo(Jiangxi Cancer Hospital,the Second Affiliated Hospital of Nanchang Medical College,Jiangxi Clinical Research Center for Cancer,Nanchang 330029,China;College of Chemistry and Bioengineering,Yichun University,Yichun 336000,China;Medical College,Yichun University,Yichun 336000,China;Jiangxi Provincial People's Hospital(the First Affiliated Hospital of Nanchang Medical College),Nanchang 330006,China)

机构地区：[1]江西省肿瘤医院,南昌医学院第二附属医院,江西省恶性肿瘤临床医学研究中心,南昌330029 [2]宜春学院化学与生物工程学院,宜春336000 [3]宜春学院医学院,宜春336000 [4]江西省人民医院(南昌医学院第一附属医院),南昌330006

出　　处：《世界科学技术-中医药现代化》2024年第9期2289-2304,共16页Modernization of Traditional Chinese Medicine and Materia Medica-World Science and Technology

基　　金：江西省教育厅科学技术研究青年项目课题(GJJ2203557):基于网络药理学和蛋白质组学探索Peiminine抗结肠癌的机制研究,负责人:何勤思;江西省主要学科学术和技术带头人培养计划——青年人才项目(20212BCJL23056):贝母素乙对炎症微环境下结肠肿瘤生物学行为的影响及其作用机制,负责人:饶军;江西省自然科学基金资助项目(20224BAB206114):基于IGF-1介导的铁死亡探讨贝母素乙逆转结肠癌化疗耐药的效应机制研究,负责人:饶军。

摘　　要：目的基于网络药理学、分子对接技术和Label-free DIA定量磷酸化蛋白组揭示贝母素乙抗结肠癌的潜在作用机制。方法①利用SwissTargetPrediction、TargetNet和Pharmmapper数据库获取贝母素乙的作用靶点,利用DisGeNET、GeneCards和OMIM数据库获取结肠癌的作用靶点,然后通过Venny2.1.0在线平台得到贝母素乙和结肠癌的交集靶点。接着运用String数据库和Cytoscape 3.8.2软件绘制交集靶点的PPI网络图并得到贝母素乙抗结肠癌的主要靶点,通过David数据库和微生信可视化云平台进行GO分析和KEGG分析。②采用MOE(Molecular operating environment)软件对贝母素乙与主要靶点进行分子对接验证。③利用Label-free DIA定量磷酸化蛋白组方法对贝母素乙处理的DT组(DT1-DT3)和对照组(NC1-NC3)的6个样本进行检测和生物学功能分析。结果①贝母素乙与结肠癌交集靶点共275个,分子对接显示贝母素乙与AKT1、EGFR、HSP90AA1和SRC的蛋白结构均能稳定对接并存在相互作用:贝母素乙与AKT1蛋白的氨基酸残基主要通过氢键发生相互作用;与EGFR、HSP90AA1和SRC蛋白的氨基酸残基主要通过离子键和氢键产生相互作用。②磷酸化蛋白组学分析显示:相对于NC组,有880个磷酸化修饰位点在DT组中显著上调(包括AKT1的S124、S126位点和EGFR的T648、S643位点),有425个磷酸化修饰位点在DT组中显著下调(包括HSP90AA1的T317位点)。③对比网络药理学和磷酸化蛋白组学分析结果发现:贝母素乙抗结肠癌的主要靶点为AKT1、EGFR和HSP90AA1,其通过调控AMPK signaling pathway、mTOR signaling pathway和Choline metabolism in cancer等17条通路促进结肠癌细胞凋亡。结论本研究揭示了贝母素乙治疗结肠癌具有多靶点、多通路的潜在机制,为后续的研究提供了一定的方向与参考。Objective Network pharmacology,Molecular docking and Label-free DIA quantitative phosphoproteomics were used to reveal the potential mechanism of Peiminine against colon cancer.Methods①The target of peiminine was obtained by SwissTargetPrediction,TargetNet and pharmmapper,and the target of colon cancer was obtained by DisGeNET,GeneCards and OMIM.Then the intersection target of Peiminine and Colon cancer was obtained by Venny2.1.0 online platform.Then,String database and Cytoscape3.8.2 software were used to map the PPI network of intersection targets,and the main targets of Peiminine against Colon cancer were obtained.GO analysis and KEGG pathway analysis were carried out through David database and Weisenxin visual cloud platform.②MOE(molecular operating environment)software was used to perform molecular docking of peiminine and the main target.③Label-free DIA quantitative phosphoproteomics was used to detect and analyze the biological function of DT group(DT1-DT3)treated with Peiminine and control group(NC1-NC3).Results①There were 275 intersection targets between peiminine and colon cancer.Molecular docking showed that peiminine could stably dock and interact with the protein structures of AKT1,EGFR,HSP90AA1 and SRC:Peiminine interacted with the amino acid residues of AKT1 mainly through hydrogen bonding.Peiminine interacted with amino acid residues of EGFR,HSP90AA1 and SRC mainly through ionic bond and hydrogen bonding.②Phosphoproteomics analysis showed that:Compared with the NC group,880 phosphorylated modification sites were significantly up-regulated in the DT group(including S124 and S126 sites of AKT1 and T648 and S643 sites of EGFR),and 425 phosphorylated modification sites were significantly down-regulated in the DT group(including T317 sites of HSP90AA1).③Comparing the results of network pharmacology and phosphoproteomics analysis,it was found that:The main targets of Peiminine against Colon cancer are AKT1,EGFR and HSP90AA1.It promotes apoptosis of colon cancer cells by regulating 17 pathway

关 键 词：贝母素乙 结肠癌 网络药理学 分子对接 磷酸化蛋白组学 

分 类 号：R285.5[医药卫生—中药学]