基于网络药理学探讨中药蟾毒灵在骨肉瘤中的抗肿瘤作用机制  

Exploring the anti-tumor mechanism of traditional Chinese medicine bufalin in osteosarcoma based on network pharmacology

作　　者：方波[1] 丁晓蕾[2] FANG Bo;DING Xiaolei(Department of Orthopaedic Surgery,Dalian Second People's Hospital,Liaoning Dalian 116011,China;Department of Oncology,Second Affiliated Hospital of Dalian Medical University,Liaoning Dalian 116023,China)

机构地区：[1]大连市第二人民医院骨外科,辽宁大连116011 [2]大连医科大学附属第二医院肿瘤科,辽宁大连116023

出　　处：《现代肿瘤医学》2024年第4期629-635,共7页Journal of Modern Oncology

基　　金：浙江省衢州市衢江区生命绿洲公益服务中心项目(编号:20211210)。

摘　　要：目的:本研究运用网络药理学分析方法,了解蟾毒灵(bufalin)在骨肉瘤中的抗肿瘤作用机制,并进一步利用相关实验进行验证。方法:通过PharmMapper数据库预测得到蟾毒灵的潜在作用靶点,借助GeneCards数据库搜索骨肉瘤相关疾病靶点,对两个数据库中的靶点进行Venny分析,随后使用String数据库绘制蛋白互作(PPI)网络,利用David数据库对关键靶点进行基因本体(gene ontology,GO)富集及KEGG信号通路分析。最后运用Cytoscape 3.7.2及Origin 2020软件构建“化合物-通路-靶点-疾病”网络图。最后利用CCK-8实验和Western blot实验进行验证。结果:通过药理学分析一共获得蟾毒灵与骨肉瘤共同靶点107个,KEGG通路富集分析共得到相关信号通路34条,涉及到肿瘤信号通路有:PI3K-AKT信号通路、MAPK信号通路、p53信号通路、VEGF信号通路、ErbB信号通路等。将其进行PPI网络分析表明,MYC、CCND1、IL6、ESR1、CDKN2A、IGF1、CAT、CDKN1A、ANXA5、PTGS2为连接度最高、权重最大的靶点。CCK-8实验证实蟾毒灵具有抑制骨肉瘤细胞增殖的作用;Western blot发现增殖相关蛋白MYC、CCND1蛋白表达下降。回复实验进一步证实,蟾毒灵可能通过调控PI3K-AKT信号通路发挥抗骨肉瘤的作用。结论:基于网络药理学分析以及相关实验发现蟾毒灵可能通过调控PI3K-AKT信号转导通路抑制骨肉瘤的生长。Objective:In this study,the network pharmacological analysis method was used to understand the anti-tumor mechanism of bufalin in osteosarcoma,and further verified by relevant experiments.Methods:The potential targets of bufalin were predicted through the PharmMapper database.The targets of osteosarcoma related diseases were searched through the GeneCards database.The targets in the two databases were analyzed with Venny.Then the String database was used to draw the protein interaction(PPI)network,and the David database was used to enrich the gene ontology(GO)and analyze the KEGG signal pathway of the key targets.Finally,Cytoscape 3.7.2 and Origin 2020 software were used to construct the network diagram of"compound-pathway-target-disease".Finally,CCK-8 experiment and Western blot experiment were used to verify.Results:A total of 107 common targets of bufalin and osteosarcoma were obtained through pharmacological analysis,and 34 related signal pathways were obtained through KEGG pathway enrichment analysis,involving PI3K-AKT signal pathway,MAPK signal pathway,p53 signal pathway,VEGF signal pathway,ErbB signal pathway,etc.The PPI network analysis showed that MYC,CCND1,IL6,ESR1,CDKN2A,IGF1,CAT,CDKN1A,ANXA5 and PTGS2 were the targets with the highest connectivity and weight.CCK-8 proved that bufalin could inhibit the proliferation of osteosarcoma cells.Western blot showed the expression of proliferation related proteins MYC and CCND1 was decreased.The recovery experiment further confirmed that bufalin may play an anti-osteosarcoma role by regulating PI3K-AKT signaling pathway.Conclusion:Based on the network pharmacological analysis and related experiments,it was found that bufalin may inhibit the growth of osteosarcoma by regulating PI3K-AKT signal transduction pathway.

关 键 词：蟾毒灵 网络药理学 骨肉瘤 抗肿瘤作用 分子机制 

分 类 号：R738.1[医药卫生—肿瘤]