Proteomic analysis of laser-microdissected paraffin-embedded tissues: (2) MRM assay for stage-related proteins upon non-metastatic lung adenocarcinoma
Proteomic analysis of laser-microdissected paraffin-embedded tissues: (2) MRM assay for stage-related proteins upon non-metastatic lung adenocarcinoma
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DOI：
10.1016/j.jprot.2009.11.010
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发表时间：
2010-04-18
影响因子：
3.3
通讯作者：
Kato, Harubumi
中科院分区：
文献类型：
--
作者：
Nishimura, Toshihide;Nomura, Masaharu;Kato, Harubumi
先前的论文通过全局鸟枪蛋白质组学和光谱计数提出了81种阶段特异性表达的IA或IIIA期蛋白质的候选物。选择6种蛋白质（这些蛋白质的子集）进行进一步验证研究，因为它们可能是可溶性和/或分泌性的，其性质便于在临床实践中检测血液中的蛋白质。多反应监测（MRM）定量分析表明，6个候选基因中napsin-A和前梯度蛋白2（hAG-2）可用于确定IA或IIIA期，并与转移有关。在这项研究中，我们注意到，具有更好结局的IIIA期患者显示出与IA期患者相似的napsin-A谱。因此，我们检查了14例额外的患者进行分析，其中包括IA期患者的较差的结果和IIIA期患者的较好的结果。所有患者的napsin-A的MRM分析表明，napsin-A含量与IA期的更好结局相关。本研究和发现研究表明，通过激光显微切割（LMD）单独直接分离肿瘤细胞大大降低了综合分析的复杂性，并且使用内源性内标的MRM质谱法是一种可行的技术，用于定量验证福尔马林固定石蜡包埋（FFPE）组织中的靶蛋白。(C)2009爱思唯尔有限公司版权所有。
A preceding paper suggested 81 candidates of stage-specifically expressed proteins for either stage IA or IIIA by global shotgun proteomics and spectral counting. Six proteins, a subset of these proteins, were chosen for a further verification study since they are potentially soluble and/or secretory, which nature is convenient for detecting them in blood in clinical practice. The multiple-reaction monitoring (MRM) quantitative analysis suggested that napsin-A and anterior gradient protein 2 homolog (hAG-2) out of the 6 candidates would be useful for determining stage IA or IIIA and are related to metastasis. In the study we noted that stage IIIA patients with better outcome showed napsin-A profiles similar to that of stage IA patients. We therefore examined 14 additional patients for analysis, which contained the IA-stage patients of poorer outcome and the IIIA-stage patients of better outcome. The MRM analysis of napsin-A for all patients suggests that napsin-A contents correlate with better outcome in stage IA. This and discovery studies demonstrate that direct isolation of tumor cells alone by laser microdissection (LMD) greatly reduces complexity on comprehensive analyses, and that MRM mass spectrometry using the endogenous internal standard is a feasible technology for quantitative verification of target proteins in formalin-fixed paraffin embedded (FFPE) tissues. (C) 2009 Elsevier B.V. All rights reserved.