目的 基于网络药理学、分子对接技术和实验验证探究狼毒大戟治疗甲状腺癌的作用机制。方法 通过SwissADME、Swiss Target Prediction数据库筛选狼毒大戟活性成分和相关作用靶点；利用GeneCards、OMIM、DisGeNET数据库筛选甲状腺癌靶点基因；基于STRING平台和Cytoscape软件构建蛋白互作网络（PPI）、狼毒大戟-有效活性成分-交集靶点网络以及筛选核心靶点；使用DAVID数据库进行基因本体（GO）功能注释分析与京都基因与基因组百科全书（KEGG）通路富集分析；利用AutoDockTools软件对狼毒大戟核心成分和关键靶点进行分子对接验证；Western blotting分析狼毒大戟对表皮生长因子受体（EGFR）/Janus激酶（JAK2）/信号转导因子和转录激活因子3（STAT3）信号通路的作用。结果 共获得狼毒大戟潜在活性成分38种、成分靶点488个，甲状腺癌靶点3 050个，交集靶点175个；狼毒大戟治疗甲状腺癌的生物过程包括磷酸化、蛋白磷酸化、细胞群增殖正调控等，相关信号通路涉EGFR/JAK2/STAT3、磷脂酰肌醇3-激酶（PI3K）/蛋白激酶B（Akt）、丝裂原活化蛋白激酶（MAPK）、EGFR、肿瘤坏死因子（TNF）、高级糖基化终末产物-受体（AGE-RAGE）、凋亡、Th17细胞分化等信号通路；分子对接结果表明狼毒大戟治疗甲状腺癌的核心成分与JAK2、EGFR、STAT3具有显著的亲和力，结合性能稳定；Western blotting实验提示岩大戟内酯A呈浓度相关性抑制EGFR/JAK2/STAT3信号通路。结论 狼毒大戟可能通过调控多个关键靶点、多种生物学过程和多条信号通路，尤其是EGFR/JAK2/STAT3信号通路发挥抗甲状腺癌效应，为今后深入研究狼毒大戟对甲状腺癌的调控机制提供理论依据。

Objective To investigate the action mechanism of Euphorbia fischeriana in treatment of thyroid cancer based on network pharmacology, molecular docking technology, and experimental validation. Methods Screening the main active ingredient and corresponding targets of E. fischeriana through databases such as SwissADME and Swiss Target Prediction. GeneCards, OMIM and DisGeNET databases were used to collect the target genes of thyroid cancer. PPI network and “E. fischeriana–active ingredient–intersection targets” network diagrams were constructed using the STRING platform and Cytoscape software, from which core targets were screened. DAVID database was applied for GO functional annotation analysis and KEGG pathway enrichment analysis, and molecular docking analysis validated binding affinity values for the core components and key targets via AutoDockTools software. Western blotting was assessed for its effects of E. fischeriana on EGFR/JAK2/STAT3 signaling pathway. Results A total of 38 potential effective constituents, 488 drug related targets, 3 050 thyroid cancer targets, and 175 overlapping targets were obtained. Biological processes mainly included phosphorylation, protein phosphorylation, and positive regulation of cell population proliferation, etc. Relevant signaling pathways involved EGFR/JAK2/STAT3, PI3K/Akt, MAPK, EGFR, TNF, AGE-RAGE, apoptosis, and Th17 cell differentiation signaling pathways. Molecular docking revealed that the core active component jolkinolide A was stable in docking with JAK2, EGFR, and STAT3, implied with significant affinity. Western blotting assay showed that E. fischeriana inhibited EGFR/JAK2/STAT3 signaling pathway in a concentration-dependent manner. Conclusion E. fischeriana can exert an antithyroid cancer effect by regulating multiple key targets, multiple biological processes and multiple pathways, especially EGFR/JAK2/STAT3 signaling pathway, which will provide theoretical reference for further in-depth investigating the regulatory mechanism of E. fischeriana on thyroid cancer.

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国家自然科学基金资助项目（82473266；82160129）；甘肃省科技重大专项（22ZD6FA054）；甘肃省创新驱动助力工程项目（GXH20230817-14）