中枢神经系统血管母细胞瘤细胞与人脑神经元细胞差异蛋白质分析  

Label-free Differential Proteomic Analysis of Central Nervous System Hemangioblastoma and Human Nerve Cells

作　　者：刘杭[1,2] 谢嵘[3] 姚鋆[2] 樊惠芝[1] 朱剑虹[3] 杨芃原[1,2] 

机构地区：[1]复旦大学化学系,上海200433 [2]复旦大学生物医学研究院,上海200032 [3]复旦大学附属华山医院神经外科,上海200040

出　　处：《分析化学》2009年第10期1439-1444,共6页Chinese Journal of Analytical Chemistry

基　　金：国家自然科学基金(No.20735005);国家重大基础研究计划(973计划;No.2007CB914100)资助

摘　　要：以人工培养的中枢神经系统血管母细胞瘤(Central nervous system hem angioblastoma,HB)细胞为研究对象,发展了蛋白质组学分析方法,鉴定了HB细胞与人脑神经元细胞的差异蛋白质。采用在线HPLC串联LTQ-Orbitrap质谱鉴定样品的可溶性蛋白质,得到了HB细胞的蛋白质组表达谱。HB细胞鉴定得到674个蛋白质,神经元细胞鉴定获得531个蛋白质。根据基于肽段鉴定的蛋白质组半定量分析方法对质谱数据进行蛋白质的差异比较分析,发现了波形蛋白(Vimentin),14-3-3epsilon蛋白和碳酸酐酶Ⅱ(Carbonic anhydraseⅡ,CAⅡ)等在HB细胞中表达量发生明显变化的蛋白质,并对其进行了免疫组织化学染色分析。结果显示,波形蛋白(Vimentin)、14-3-3epsilon蛋白以及碳酸酐酶Ⅱ(CAⅡ)等蛋白质表达量的改变与HB的发病密切相关,对探索HB的起源有重要意义。The proteomes of central nervous system hemangioblastoma（CNS HB） cells and human nerve cells were analyzed with online HPLC in combination with LTQ-Orbitrap mass spectrometry.After database searching,647 proteins of HB cells,meanwhile 531 proteins of human nerve cells were identified.A label-free technique based on half-quantitative analysis was performed between those two proteomes.The results showed that the expression levels of Vimentin,14-3-3 epsilon protein,carbonic anhydrase Ⅱ were significantly changed in HB cells.The proteome results were also validated with immunohistochemical staining.The results demonstrate this method has important implications for the identifying differences in proteomic profiles of HB samples and the design biomarker studies for CNS HB.

关 键 词：中枢神经系统血管母细胞瘤 蛋白质组学 高效液相色谱-质谱 半定量分析 

分 类 号：R739.4[医药卫生—肿瘤]