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首页> 美国卫生研究院文献>Journal of Histochemistry and Cytochemistry >Quantitative Proteomic Analysis of Formalin-fixed and Paraffin-embedded Nasopharyngeal Carcinoma Using iTRAQ Labeling Two-dimensional Liquid Chromatography and Tandem Mass Spectrometry
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Quantitative Proteomic Analysis of Formalin-fixed and Paraffin-embedded Nasopharyngeal Carcinoma Using iTRAQ Labeling Two-dimensional Liquid Chromatography and Tandem Mass Spectrometry

机译：使用iTRAQ标记二维液相色谱和串联质谱对福尔马林固定和石蜡包埋的鼻咽癌进行蛋白质组学定量分析

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Formalin-fixed, paraffin-embedded (FFPE) tissue specimens represent a potentially valuable resource for protein biomarker investigations. In this study, proteins were extracted by a heat-induced antigen retrieval technique combined with a retrieval solution containing 2% SDS from FFPE tissues of normal nasopharyngeal epithelial tissues (NNET) and three histological types of nasopharyngeal carcinoma (NPC) with diverse differentiation degrees. Then two-dimensional liquid chromatography-tandem mass spectrometry coupled with isobaric tags for relative and absolute quantification (iTRAQ) labeling was employed to quantitatively identify the differentially expressed proteins among the types of NPC FFPE tissues. Our study resulted in the identification of 730 unique proteins, the distributions of subcellular localizations and molecular functions of which were similar to those of the proteomic database of human NPC and NNET that we had set up based on the frozen tissues. Additionally, the relative expression levels of cathepsin D, keratin8, SFN, and stathmin1 identified and quantified in this report were consistent with the immunohistochemistry results acquired in our previous study. In conclusion, we have developed an effective approach to identifying protein changes in FFPE NPC tissues utilizing iTRAQ technology in conjunction with an economical and easily accessible sample preparation method. >(J Histochem Cytochem 58:517–527, 2010)
机译：福尔马林固定，石蜡包埋（FFPE）的组织标本代表了蛋白质生物标志物研究的潜在宝贵资源。在这项研究中，蛋白质是通过热诱导抗原提取技术与含有2％SDS的提取液相结合，从正常鼻咽上皮组织（NNET）的FFPE组织和三种不同分化程度的鼻咽癌（NPC）组织学类型中提取的。然后采用二维液相色谱-串联质谱联用等压标记进行相对和绝对定量（iTRAQ）标记，以定量鉴定NPC FFPE组织类型中差异表达的蛋白质。我们的研究结果鉴定出730种独特的蛋白质，其亚细胞定位的分布和分子功能与我们基于冷冻组织建立的人类NPC和NNET的蛋白质组数据库相似。此外，在本报告中鉴定和定量的组织蛋白酶D，角蛋白8，SFN和stathmin1的相对表达水平与我们先前研究中获得的免疫组织化学结果一致。总之，我们已经开发出一种有效的方法，可以利用iTRAQ技术结合经济且易于获取的样品制备方法来鉴定FFPE NPC组织中的蛋白质变化。 >（J Histochem Cytochem 58：517-527，2010）

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