中枢神经系统血管母细胞瘤的蛋白质组学分析

Proteomics Analysis of Central Nervous System Hemangioblastomas

背景与目的:中枢神经系统血管母细胞(centralnervoussystemhemangioblastoma,CNSHB)的治疗较难,至今病因不明。本研究应用蛋白质组学方法分析HB与正常脑组织的蛋白质表达谱的差异,分离鉴定与HB发病的蛋白质,为研究HB的组织学起源与发病机制提供依据。方法:收集5例血管母细胞瘤与5例正常脑组织标本。提取标本的总蛋白质。通过双向凝胶电泳(2-DE)分离蛋白,确定血管母细胞瘤与正常脑组织的差异表达蛋白质点。应用基质辅助激光解离-飞行时间质谱(MALDI-TOF-MS)鉴定并对鉴定蛋白质进行生物信息学分析。结果:通过双向凝胶电泳,建立了约含600个蛋白质点的血管母细胞瘤及正常脑组织的高分辨率蛋白质表达图谱。两者相比具有较高的同源性。应用ImageMaster2D图像分析软件共发现115个差异蛋白质点。经MALDI-TOF-MS分析后成功鉴定了87个蛋白质点共计47种蛋白质。其中HB组较正常对照组表达上调46个蛋白质点,下调41个蛋白质点。鉴定蛋白质的功能涉及转运、蛋白质折叠与代谢、三羧酸循环、细胞分化、干细胞相关蛋白质等数个方面。对鉴定所得蛋白质Vimentin、14-3-3蛋白进行免疫组化染色可重复本研究的结果。结论:中枢神经系统血管母细胞瘤可能是一种起源于间叶脑组织的肿瘤。其发生是一个多因子参与、多步骤的复杂过程。多种蛋白质如Vimentin及14-3-3蛋白参与了血管母细胞瘤的发病并起到重要的作用。

BACKGROUND ＆ OBJECTIVE：To establish detailed protein expression maps of central nervous system hemangioblastomas {CNS HB） and identify discrepant proteins between HB and normal brain tissues by using proteomics analysis so as to better understand the histological origin of CNS HB. METHODS ： Ten individual brain regions with five HB tissues （HB group） and five normal cerebellar tissues （control group） were included. Total proteins were extracted from the samples and process of two dimensional gel electrophoresis, spot excision, in-gel digestion, MALDI-TOF MS and bioinformatics analysis were carried out. RESULT： After 2-DE, high-resolution maps containing about 600 protein spots of HB and normal brain tissues were produced. The 2-DE maps show a satisfactory homology between HB and normal brain tissues. By using the ImageMaster 2D software, 115 discrepant protein spots between HB and normal brain tissues could be selected and 87 of them were successfully identified by MALDI-TOF MS. Among them, 46 proteins expressions were upregulated while 41 were down-regulated. According to their functional role, the identified proteins could be divided into several groups including transport, protein metabolism and folding, glycolysis and stem cell proteins. The results of immunohistochemistry staining of Vimentin and 14-3-3 proteins could authenticate the validity of the proteomics analysis in this study. CONCLUSIONS： The process of CNS HB occurring is an intricate, mulficomponent, mulfifactorial and multi-step process which is mediated by a variety of proteins. CNS HB as a kind of tumors may come from normal brain mesenchymal tissues. Several proteins such as Vimentin and 14-3-3 may play an important role in the occurring of HB.