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目的:探究胃肠神经内分泌肿瘤（GI-NENs）原发灶与相应瘤旁组织及肝转移灶中的蛋白表达差异。方法:选取2015年7月至2019年4月于中国医学科学院肿瘤医院手术治疗的9例GI-NENs伴肝脏转移患者的原发灶、肝转移灶组织及相应瘤旁组织，采用数据非依赖性采集（DIA）技术检测其蛋白表达，以 P＜0.05且|log 2FC|＞0.5（FC为差异倍数）作为判定标准，明确原发灶与相应瘤旁组织、原发灶与肝转移灶、不同分化程度原发灶、不同分化程度肝转移灶的差异表达蛋白。对差异表达蛋白进行火山图分析、聚类分析、基因本体（GO）功能分析和京都基因与基因组百科全书（KEGG）通路富集分析。 结果:相对于瘤旁组织，胃NENs原发灶中有85种蛋白表达下调，42种蛋白表达上调，差异表达蛋白主要富集在三磷酸鸟苷酶活性调控和脱氧核糖核苷单磷酸盐分解代谢相关的生物过程、糖胺聚糖生物合成-硫酸软骨素/硫酸皮肤素和泛酸盐与CoA生物合成信号通路。肠NENs原发灶中有114种蛋白表达下调，155种蛋白表达上调，差异表达蛋白主要富集在谷胱甘肽代谢和硫化合物代谢相关的生物过程、收集导管酸分泌和牛磺酸、次牛磺酸代谢信号通路。相对于神经内分泌癌（NECs）原发灶，G1～2分化原发灶中有168种蛋白表达下调，278种蛋白上调，差异表达蛋白显著富集在DNA代谢和DNA复制的生物学过程，以及复制和错配修复等通路。相对于NECs转移灶，G1～2分化转移灶中有95种蛋白表达下调，97种蛋白表达上调，差异表达蛋白显著富集到转录共激活子的活性和催化活性功能、碱基切除修复和蛋白外排通路。相对于G1分化的原发灶，G1分化的转移灶中有530种蛋白表达下调，211种蛋白表达上调。相对于G2分化的原发灶，G2分化的转移灶中有53种蛋白表达下调，96种蛋白表达上调。相对于NECs原发灶，NECs转移灶中有109种蛋白表达下调，92种蛋白表达上调。G1和G2分化的GI-NENs原发灶与转移灶差异表达蛋白富集的信号通路中有多条相似，而GI-NECs原发灶与转移灶差异表达蛋白富集的信号通路中只有1条与GI-NENs原发灶与转移灶差异表达蛋白富集的信号通路相同，即药物代谢信号通路。G1分化的原发灶与转移灶差异表达蛋白主要表达于细胞质（20.26%）、线粒体（18.67%）和细胞核（15.48%）。G2分化的原发灶与转移灶差异表达蛋白主要表达于细胞质（20.24%）、细胞核（18.25%）和细胞膜（15.08%）。NECs原发灶与转移灶差异表达蛋白主要表达于细胞核（23.78%）、细胞质（22.7%）和细胞膜（11.35%）。结论:不同部位和分化程度的GI-NENs原发灶、瘤旁组织及转移灶中蛋白表达差异明显。

Objective:To investigate the differences of protein expressions in the primary tumors, adjacent tissues, and metastatic tumors of gastrointestinal neuroendocrine neoplasms.Methods:Nine patients with gastrointestinal neuroendocrine tumors (GI-NENs) with liver metastasis who underwent surgery at the National Cancer Center/Cancer Hospital, Chinese Academy of Medical Sciences from July 2015 to April 2019 were selected. The protein expressions of the primary tissues, liver metastatic tissues, and adjacent tissues were detected by the data independent acquisition (DIA) technology. P<0.05 and | log 2FC|>0.5 (FC as the difference multiple) were used as the criteria to identify the differentially expressed proteins in the primary tissues vs adjacent tissues, primary tissues vs liver metastatic tissues, primary tissues with different degrees of differentiation, and liver metastatic tissues with different degrees of differentiation. The differentially expressed proteins were investigated by volcano map analysis, cluster analysis, Gene Ontology (GO) function analysis, and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway enrichment analysis. Results:Compared with adjacent tissues, 85 proteins were downregulated and 42 proteins were upregulated in the primary tissues of gastric NENs. The differentially expressed proteins were mainly enriched in the biological processes related to the regulation of guanosidase triphosphate activity and the catabolism of deoxyribonucleoside monophosphate, the glycosaminoglycan biosynthesis chondroitin sulfate/dermatan sulfate, pantothenate, and CoA biosynthesis signaling pathways. 114 proteins were downregulated and 155 proteins were upregulated in the primary tissues of intestinal NENs. The differentially expressed proteins were mainly enriched in the biological processes related to glutathione metabolism and sulfur compound metabolism, collecting duct acid secretion, and taurine and hytaurine metabolism signaling pathways. Compared with the primary tissues of neuroendocrine cancers (NECs), 168 proteins were downregulated and 278 proteins were upregulated in G1-2 differentiation primary tissues. The differentially expressed proteins were significantly enriched in biological processes such as DNA metabolism and DNA replication, as well as replication, mismatch repair, and other pathways. Compared with the metastatic tissues of NECs, 95 proteins were downregulated and 97 proteins were upregulated in G1-2 differentiated metastases. The differentially expressed proteins were significantly enriched in the activity and catalytic activity of transcriptional coactivators, base excision repair, and protein efflux pathways. Compared with G1 differentiated primary tissues, 530 proteins were downregulated and 211 proteins were upregulated in G1 differentiated metastatic tissues. Compared with G2 differentiated primary lesions, 53 proteins were downregulated and 96 proteins were upregulated in G2 differentiated metastatic tissues. Compared with the primary lesions of NECs, 109 proteins were downregulated and 92 proteins were upregulated in the metastatic tissues of NECs. In G1 and G2 differentiated GI-NENs, there are many similar signal pathways enriched in differentially expressed proteins between primary lesions and metastases, while only one signal pathway enriched in differentially expressed proteins between primary and metastatic tissues of NECs is the same as that enriched in differentially expressed proteins between primary and metastatic tissues of GI-NENs, which is the drug metabolism signal pathway. The differentially expressed proteins in G1 differentiated primary and metastatic tissues were mainly expressed in cytoplasm (20.26%), mitochondria (18.67%), and nucleus (15.48%). The differentially expressed proteins in the primary and metastatic tissues of G2 differentiation were mainly expressed in the cytoplasm (20.24%), nucleus (18.25%), and cell membrane (15.08%). The differentially expressed proteins in the primary and metastatic tissues of NECs were mainly expressed in the nucleus (23.78%), cytoplasm (22.7%), and cell membrane (11.35%).Conclusion:The protein expressions of GI-NENs in the primary tissues, adjacent tissues, and metastatic tissues were significantly different in different sites and degrees of differentiation.