肾母细胞瘤血清非应激相关性蛋白质标志物的筛选及鉴定  

Detection and identification of non - stress protein biomarkers for nephroblastoma

作　　者：王磊[1] 范应中[1] 张谦[1] 王家祥[1] 

机构地区：[1]郑州大学第一附属医院小儿外科,450052

出　　处：《中华实用儿科临床杂志》2016年第11期836-839,共4页Chinese Journal of Applied Clinical Pediatrics

基　　金：国家自然科学基金（81172085）

摘　　要：目的通过蛋白质组学技术检测肾母细胞瘤患儿血清与健康儿童血清中的差异性蛋白质，筛选出。肾母细胞瘤非应激相关性蛋白质标志物。方法收集2010年5月至2014年5月郑州大学第一附属医院接诊的。肾母细胞瘤患儿、健康体检儿童及外伤1～3h患儿血清。通过蛋白质组学的表面增强激光解吸电离飞行时间质谱（SELDI-TOF—MS）、基质辅助激光解吸电离飞行时间质谱（MALDI-TOF-MS）、高效液相色谱（HPLC）以及二维液相色谱一线性离子阱质谱（2D-LC—LTQ—MS）相关技术，对血清差异性蛋白质进行筛选并排除创伤应激蛋白质干扰；鉴定肾母细胞瘤血清非应激相关性蛋白质标志物。结果SELDI—TOF—MS筛选出质荷比为6630．58Da的蛋白质或肽段为特异性标志物，对比确认创伤应激患儿血清质谱中无相同或相似蛋白质。通过HPLC纯化及MALDI—TOF—MS和2D-LC-LTQ-MS鉴定，发现肾母细胞瘤患儿血清非应激相关性蛋白质为载脂蛋白CI（APOCI）。结论在排除应激干扰因素下，通过蛋白质组学相关技术筛选并鉴定出APOCI为一种肾母细胞瘤患儿血清蛋白质标志物，为进一步研究肾母细胞瘤患儿血清中蛋白质表达水平改变的机制及早期诊断肾母细胞瘤提供了可能。Objective To test the differential proteomics by using proteomic technology of children diagnosed with nephroblastoma and healthy children,thereby for screening non - stress - related protein markers of Wilms＇ tumor. Methods The serum of children with Wilms＇ tumor, children with trauma in 1 - 3 hours and healthy children were collected in the First Affiliated Hospital of Zhengzhou University from May 2010 to May 2014. Then, the differential pro- teomics were screened and the interference of traumatic stress proteins in the process were eliminated by using proteomic technology of surfaced enhanced laser desorption/ionization time of flight mass spectroscopy （ SELDI - TOF - MS）, matrix - assisted laser desorption/ionization time - of - flight mass spectroscopy （ MALDI - TOF - MS）, high per- formance liquid chromatography（ HPLC）, and two dimensions -liquid chromatography- linear trap quadrupole -mass spectrometer（2D -LC -LTQ -MS）, therefore the non -stress -related protein biomarkers of Wilms＇ tumor were de- termined. Results A mass - to - charge ratio 6 630.58 Da protein or peptide was selected as tumor - specific marker. And there were no same or similar proteins in mass spectrometry of children with traumatic stress by SELDI - TOF - MS. Finally, by purification through HPLC and identification through MALDI - TOF - MS and 2D - LC - LTQ - MS, apolipoprotein CI（ APO CI） was found to be the non - stress - related serum protein of Wilms＇ tumor. Conclusions It is identified that APO CI is an important serum protein biomarker of Wilms＇ tumor by using proteomic technology in eliminating the influences of interference factors of stress. Therefore, the results provide possibilities of further studies and investigating the mechanisms of the protein expression changes and early diagnosing the Wilms＇ tumor.

关 键 词：蛋白质组学 肾母细胞瘤 蛋白质标志物 应激 

分 类 号：R737.11[医药卫生—肿瘤]