应用iTRAQ技术筛选DNT与低级别胶质瘤差异表达蛋白

发布时间：2018-03-01 08:06

本文关键词： 胚胎发育不良性神经上皮肿瘤低级别胶质瘤蛋白质组学同位素标记相对和绝对定量技术生物标记物　出处：《中华全科医学》2016年07期 　论文类型：期刊论文

【摘要】：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P0.05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。
[Abstract]:Objective to screen differentially expressed proteins between embryonic dysplasia neuroepithelial tumors and low-grade gliomas by isotope labeling and absolute quantitative proteomics combined with liquid phase tandem mass spectrometry. Methods embryonic development was collected. Frozen fresh specimens were collected from 6 cases of neuroepithelial tumors (No. 113) and 6 cases of low-grade gliomas (No. 114), respectively. The samples of each group were mixed and separated by protein extraction, protein concentration measurement (Bradford quantitative analysis, polyacrylamide gel electrophoresis, protein enzymolysis iTRAQ labeled SCX, and then analyzed by liquid / mass spectrometry based on QE). Using protein identification software Mascot 2.3.02, Uni Prot-Human database was selected, and then database search and bioinformatics analysis were carried out. Results 88 differentially expressed proteins of DNT were identified, including 44 up-regulated proteins and 44 down-regulated proteins, which had different biological activities. Important proteins include water channel, serine / threonine kinase, intracellular chloride channel protein 1, integrin A 1, glutamine synthetase, and so on. Chondroitin sulfate polysaccharide protein 4, S100A9, S100A16, S100A13, isocitrate dehydrogenase, etc. Conclusion iTRAQ technique is practical and reliable, and can effectively screen differentially expressed proteins between embryonic dysplasia neuroepithelial tumors and low-grade gliomas.
【作者单位】：广州军区广州总医院病理科;广州军区广州总医院医学实验科;
【基金】：广东省科技计划项目(2012B031800177)
【分类号】：R739.41

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