目的 基于网络毒理学、分子对接和分子动力学模拟探讨邻苯二甲酸二(2-乙基己基)酯（DEHP）与非小细胞肺癌相关的分子靶点及作用机制。方法 整合GEO数据库中非小细胞肺癌相关转录组数据，通过差异表达分析和加权基因共表达网络分析（WGCNA）筛选疾病相关基因。结合ChEMBL、SwissTargetPrediction、SEA数据库获取DEHP候选靶点，开展蛋白相互作用网络（PPI）分析、基因本体论（GO）功能及京都基因和基因组百科全书（KEGG）通路富集分析，并采用分子对接、100 ns分子动力学模拟及分子力学-泊松-玻尔兹曼表面积（MM-PBSA）结合自由能计算评估候选蛋白与DEHP的相互作用。结果 共筛选获得5 136个差异表达基因及138个DEHP候选靶点，经交集分析获得54个DEHP与非小细胞肺癌的潜在关联靶点。PPI网络显示基质金属蛋白酶（MMP）9、MMP2、信号传导及转录激活蛋白1-（STAT1）等蛋白处于网络核心位置。GO功能及KEGG通路富集分析提示，相关靶点主要涉及细胞外基质重塑、蛋白水解及相关信号转导等生物学过程。分子对接结果显示，DEHP与多个候选蛋白具有一定结合潜力，其中MMP9、MMP7与DEHP的预测结合能分别为-8.0、-6.0 kcal/mol（1 cal＝4.2 J）。分子动力学模拟显示，MMP7-DEHP和MMP9-DEHP复合物在100 ns模拟过程中均表现出相对稳定的构象特征。MM-PBSA分析结果显示，2种复合物的结合自由能分别为-136.38、-109.01 kJ/mol，范德华作用和疏水相互作用可能是维持复合物稳定的主要因素。结论 DEHP可能通过多靶点、多通路网络参与非小细胞肺癌相关生物学过程，其中MMP家族介导的细胞外基质降解和组织重塑可能是重要方向。MMP7和MMP9可作为进一步探索相关分子机制的候选靶点。

Objective To investigate the molecular targets and mechanisms linking di-(2-ethylhexyl) phthalate (DEHP) to non-small cell lung cancer based on network toxicology, molecular docking, and molecular dynamics simulation. Methods non-small cell lung cancer-related transcriptomic data from the GEO database were integrated, and disease-related genes were screened using differential expression analysis and weighted gene co-expression network analysis (WGCNA). DEHP-related candidate targets were obtained from the ChEMBL, SwissTargetPrediction, and SEA databases. PPI network analysis, GO functional enrichment, and KEGG pathway enrichment analyses were performed. Molecular docking, 100-ns molecular dynamics simulation, and MM-PBSA binding free-energy calculation were further used to evaluate the potential interactions between candidate proteins and DEHP. Results A total of 5 136 differentially expressed genes and 138 DEHP candidate targets were identified, and 54 potential DEHP- non-small cell lung cancer-associated targets were obtained by intersection analysis. The PPI network showed that MMP9, MMP2, STAT1, and other proteins occupied central positions in the network. GO functional and KEGG pathway enrichment analyses suggested that these targets were mainly involved in biological processes such as extracellular matrix remodeling, proteolysis, and related signal transduction. Molecular docking showed that DEHP had potential binding affinity with several candidate proteins, among which the predicted binding energies of MMP9 and MMP7 with DEHP were -8.0 and -6.0 kcal/mol (1 cal = 4.2 J), respectively. Molecular dynamics simulation showed that both MMP7-DEHP and MMP9-DEHP complexes exhibited relatively stable conformational characteristics during the 100-ns simulation. MM-PBSA analysis showed that the binding free energies of the two complexes were -136.38 and -109.01 kJ/mol, respectively, suggesting that van der Waals and hydrophobic interactions may be the main factors maintaining complex stability. Conclusion DEHP may participate in non-small cell lung cancer-related biological processes through a multi-target and multi-pathway network, in which MMP-mediated extracellular matrix degradation and tissue remodeling may represent important mechanisms. MMP7 and MMP9 may serve as candidate targets for further exploration of related molecular mechanisms.

R965

内蒙古自治区自然基金项目（2024MS08067）；内蒙古自治区本级事业单位引进人才科研启动支持项目（RCQD19003）；内蒙古自治区研究生科研创新项目（KC2025074S）