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目的应用同位素标记相对和绝对定量蛋白质组学技术(iTRAQ)联合液相串联质谱筛选胚胎发育不良性神经上皮肿瘤与低级别胶质瘤的差异表达蛋白。方法收集胚胎发育不良性神经上皮肿瘤(编号113)与低级别胶质瘤(编号114)各6例实体组织冻存新鲜标本,各组标本混合,通过蛋白质提取,蛋白质浓度测量(采用Bradford定量),聚丙烯酰胺凝胶电泳,蛋白质酶解,iTRAQ标记,SCX分离,再进行基于QE的液质联用分析,得到信息数据。使用蛋白质鉴定软件Mascot 2.3.02,选择Uni Prot-Human数据库,然后进行数据库搜索和生物信息学分析。依据蛋白质丰度水平,当差异倍数达到1.3倍以上,且经统计检验其P<0.05时,视为差异蛋白。结果鉴定出了中国大陆黄种人DNT相对于低级别胶质瘤的差异蛋白质88个,其中上调蛋白质44个,下调蛋白质44个。这些差异蛋白具有不同生物学活性,并参与多种代谢及信号通路。其中重要的蛋白有水通道膜内在蛋白1、丝氨酸/苏氨酸激酶、细胞内氯离子通道蛋白1、膜联蛋白A1、谷氨酰胺合成酶、硫酸软骨素多糖蛋白4、S100A9、S100A16、S100A13、异柠檬酸脱氢酶等。结论 iTRAQ技术实用可靠,有效筛选出胚胎发育不良性神经上皮肿瘤与低级别胶质瘤的差异表达蛋白。
Objective To screen differentially expressed proteins between embryonic dysplastic neoplasms and low-grade glioma using isotope labeled relative and absolute quantitative proteomics (iTRAQ) combined with liquid chromatography tandem mass spectrometry. Methods Fresh tissue samples from 6 tissues of embryonic dysplastic neuroepithelial neoplasia (n ° 113) and low grade gliomas (n = 114) were collected and the samples of each group were mixed and extracted. Protein concentration and protein concentration (determined by Bradford) , Polyacrylamide gel electrophoresis, proteolysis, iTRAQ labeling, SCX separation, and then based on QE LC / MS analysis, to obtain information data. Using Mascot 2.3.02, a protein identification software, the Uni Prot-Human database was selected for database search and bioinformatics analysis. According to the level of protein abundance, when the difference multiple reached 1.3 times or more, and the statistical test of P <0.05, as a differential protein. Results Eighty-eight differential proteins of DNT in Chinese mainland were identified, including 44 up-regulated proteins and 44 down-regulated proteins. These differential proteins have different biological activities and are involved in various metabolic and signaling pathways. Important proteins are water channel membrane intrinsic protein 1, serine / threonine kinase, intracellular chloride channel 1, annexin A1, glutamine synthetase, chondroitin sulfate 4, S100A9, S100A16, S100A13 , Isocitrate dehydrogenase and the like. Conclusion The iTRAQ technique is practical and reliable, and effectively differentiates differentially expressed proteins between embryonic dysplastic neuroepithelial tumors and low-grade gliomas.