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应用iTRAQ技术筛选儿童皮质发育障碍性癫痫差异表达蛋白

, PP. 1400-1405

Keywords: 皮质发育障碍,癫痫,蛋白质组学,iTRAQ,儿童

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Abstract:

目的筛选、鉴定儿童皮质发育障碍性癫痫的差异表达蛋白。方法以儿童皮质发育障碍性癫痫脑组织、脑外伤术后的儿童正常脑组织(对照组)为对象,采用同位素相对标记与绝对定量技术(iTRAQ)、双向液相色谱与串联质谱(2D-LC-MS/MS)鉴定儿童皮质发育障碍性癫痫组织中差异表达蛋白,对所获得差异蛋白进行生物信息学分析,筛选出具有潜在研究价值的候选蛋白,最后应用实时定量PCR和Westernblot验证这些候选蛋白在儿童皮质发育障碍性癫痫中的表达。结果共鉴定出153种差异表达蛋白(64种蛋白上调,89种蛋白下调)。Geneontology(GO)功能分析提示这些差异性蛋白主要参与酶催化活性的调节,蛋白质功能相互作用网状图发现转铁蛋白(transferrin,TF)、载脂蛋白A1(apolipoproteinA1,APOA1)、α2-巨球蛋白(alpha-2-macroglobulin,A2M)、三磷酸腺苷酶(ATPase,ATP6V1A)、波形蛋白(vimentin,VIM)处于网状图的交叉点。运用实时定量PCR及Westernblot验证,发现这5种蛋白质mRNA及蛋白表达水平与质谱结果一致(P<0.05)。结论采用iTRAQ结合2D-LC-MS/MS技术,能有效地筛选出儿童皮质发育障碍性癫痫中差异表达的蛋白。

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