基于质谱技术对垂体嗜酸性细胞瘤异常代谢通路的研究  

The abnormal metabolic pathway in the pituitary oncocytomas based on mass spectrometry

作　　者：杨晶晶 冯洁[1] 周洋 洪礼传[1] 于升远 徐宁 许国旺[2] 张亚卓[1] Yang Jingjing;Feng Jie;Zhou Yang;Hong Lichuan;Yu Shengyuan;Xu Ning;Xu Guowang;Zhang Yazhuo(Beijing Neurosurgical Institute, Capital Medical University, Beijing 100070, China;CAS Key Laboratory of Separation Science for Analytical Chemistry, Dalian Institute of Chemical Physics, Chinese Academy of Sciences, Dalian 116023, China)

机构地区：[1]首都医科大学,北京市神经外科研究所,100070 [2]中国科学院大连化学物理研究所,116023

出　　处：《中华神经外科杂志》2019年第4期395-399,共5页Chinese Journal of Neurosurgery

基　　金：国家自然科学基金(81702455)。

摘　　要：目的基于质谱技术探寻垂体嗜酸性细胞瘤的异常代谢通路。方法本研究取材于首都医科大学附属北京天坛医院神经外科行手术治疗的25例垂体嗜酸性细胞瘤患者,正常垂体组织取材于18例交通意外后尸体捐献者。采用超高效液相色谱和蛋白质质谱分析垂体嗜酸性细胞瘤与正常垂体之间的差异蛋白质。采用代谢组学分析垂体嗜酸性细胞瘤与正常垂体之间的差异代谢物。进一步通过生物信息学分析方法识别与垂体嗜酸性细胞瘤相关的信号通路。采用免疫印迹法验证c-Myc蛋白在垂体嗜酸性细胞瘤和正常垂体组织中的表达情况。结果蛋白质质谱分析结果显示,598个蛋白质在垂体嗜酸性细胞瘤和正常垂体中的表达差异均有统计学意义(均P<0.05)。代谢组学分析结果显示,95个代谢物在垂体嗜酸性细胞瘤和正常垂体中的表达差异均有统计学意义(均P<0.05)。整合通路生物信息学方法分析结果显示,10个最明显的富集通路包括代谢通路、免疫系统通路、固有免疫系统通路、蛋白质代谢通路、中性粒细胞脱颗粒通路、翻译后蛋白修饰通路、囊泡介导的转运通路、膜运输通路、止血通路及EGFR1通路。生物信息学方法分析结果显示,10个最明显的富集通路包括氧化磷酸化通路、c-Myc靶点通路1、脂肪酸代谢通路、脂肪酸合成通路、mTORC1信号通路、上皮间质转化通路、缺氧通路、糖酵解通路、外源性物质代谢通路和血红蛋白代谢通路。免疫印迹法分析结果显示,c-Myc蛋白在垂体嗜酸性细胞瘤中的相对表达量(1.58)显著高于在正常垂体中的相对表达量(0.56)(P<0.05)。结论垂体嗜酸性细胞瘤中存在异常代谢通路,且调节异常代谢的重要蛋白c-Myc表达升高。Objective To identify the abnormal metabolic pathway of pituitary oncocytoma based on Mass spectrum. Methods In this study, all 25 tumor samples were obtained following trans-sphenoidal surgery performed at Department of Neurosurgery, Beijing Tiantan Hospital, Capital Medical University. All 18 healthy pituitary glands were used as controls and obtained from donors who died from accidents. Proteomics analysis was used to analyze the differentiated expressed proteins between between pituitary oncocytoma and normal pituitary glands by nano liquid chromatography tandem mass spectrometry (nanoLC-MS/MS). Metabolomics analysis were performed to analyze the differentiated metabolites between pituitary oncocytoma and normal pituitary glands. The bioinformatics methods were used for analyzing the abnormal metabolic pathways related to pituitary oncocytoma. The expression of c-Myc was validated by western-blot method among pituitary oncocytoma and normal pituitary glands. Results Proteomics analysis showed that a total of 598 proteins were significantly different expression between pituitary oncocytoma and normal pituitary glands (all P<0.05). Metabolomics analysis displayed 95 metabolites with significant differences between pituitary oncocytoma and normal pituitary glands (all P<0.05). Ten of the most obvious enrichment pathways were shown by the Integrated Molecular Pathway Level Analysis of differentially expressed proteins and metabolites, including metabolism, immune System, innate immune system, metabolism of proteins, neutrophil degranulation, post-translational protein modification, vesicle-mediated transport, membrane trafficking, hemostasis and EGFR1 pathway. Ten of the most obvious enrichment pathways were shown by gene set enrichment analysis of differentially expressed proteins, including HALLMARK_OXIDATIVE_PHOSPHORYLATION, HALLMARK_MYC_TARGETS_V1, HALLMARK_FATTY_ACID_METABOLISM, HALLMARK_ ADIPOGENESIS, HALLMARK_MTORC1_SIGNALING, HALLMARK_EPITHELIAL_ MESENCHYMAL_TRANSITION, HALLMARK_HYPOXIA HALLMARK_GLYCOLYSI

关 键 词：垂体肿瘤 蛋白质组学 代谢组学 质谱分析法 嗜酸性细胞瘤 

分 类 号：R736.4[医药卫生—肿瘤]