Discovery of potential colorectal cancer serum biomarkers through quantitative proteomics on the colonic tissue interstitial fluids from the AOM-DSS mouse model.
Discovery of potential colorectal cancer serum biomarkers through quantitative proteomics on the colonic tissue interstitial fluids from the AOM-DSS mouse model.
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DOI：
10.1016/j.jprot.2015.11.013
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发表时间：
2016-01
影响因子：
3.3
通讯作者：
Yang Wang;Q. Shan;Guixue Hou;Ju Zhang;J. Bai;Xiaolei Lv;Yingying Xie;Huishan Zhu;Siyuan Su
中科院分区：
文献类型：
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作者：
Yang Wang;Q. Shan;Guixue Hou;Ju Zhang;J. Bai;Xiaolei Lv;Yingying Xie;Huishan Zhu;Siyuan Su
使用iTRAQ进行定量蛋白质组学分析，以发现组织间质液（TIF）中的结直肠癌（CRC）相关蛋白。使用氧化偶氮甲烷-葡聚糖硫酸钠（AOM-DSS）产生典型的炎症相关CRC小鼠模型，并且在CRC发展期间的四个阶段从这些小鼠收集TIF。使用严格的标准，共有144种蛋白质在肿瘤生长过程中显示出丰度的变化，其中45种连续增加，17种连续减少，82种不规则变化。在这144种蛋白质中，24种连续变化的蛋白质在单个TIF样品中使用MRM测量，并且18种被验证。使用MRM检查了12种经验证在TIF中连续增加的蛋白质，以评价其在个体小鼠血清样品中丰度的变化。富含亮氨酸的α-2-糖蛋白1（LRG 1）、微管蛋白β-5链（TUBB 5）和免疫球蛋白J链（IGJ）在CRC小鼠中的丰度显著高于对照小鼠。使用临床样本和MRM，我们进一步验证了LRG 1和TUBB 5是潜在的CRC血清生物标志物。这些数据表明，在动物模型中将动态TIF蛋白质组学与靶向血清蛋白质组学相结合是一种有前途的途径，用于寻求发现肿瘤血清生物标志物。然而，在临床实践中，很少有CRC生物标志物具有令人满意的特异性和敏感性。探索更多的CRC生物标志物，特别是血清中的生物标志物，是CRC研究中的一项紧迫而耗时的活动。我们的研究表明，定量评价AOM-DSS小鼠结肠组织间质液中的时相依赖蛋白是探索CRC相关蛋白和潜在血清生物标志物的可行且有效的方法。我们鉴定了两种蛋白质，LRG 1和TUBB 5，它们可能在人类临床样品中作为CRC血清生物标志物是可行的。综上所述，本研究为探索肿瘤发生的关键因素提供了一个新的角度，为潜在的血清生物标志物的发现和验证提供了新的途径。
Quantitative proteomic analysis was performed using iTRAQ to discover colorectal cancer (CRC)-related proteins in tissue interstitial fluids (TIFs). A typical inflammation-related CRC mouse model was generated using azoxymethane–dextran sodium sulfate (AOM–DSS), and TIFs were collected from these mice in four stages during CRC development. Using stringent criteria, a total of 144 proteins displayed changes in their abundances during tumor growth, including 45 that consecutively increased, 17 that consecutively decreased and 82 that changed irregularly. Of these 144 proteins, 24 of the consecutively changed proteins were measured using MRM in individual TIF samples, and 18 were verified. Twelve proteins verified to be consecutively increased in TIFs were examined using MRM to evaluate changes in their abundance in individual mouse serum samples. The abundances of leucine-rich alpha-2-glycoprotein 1 (LRG1), tubulin beta-5 chain (TUBB5) and immunoglobulin J chain (IGJ) were significantly higher in CRC mice than in control mice. Using clinical samples and MRM, we further verified that LRG1 and TUBB5 are potential CRC serum biomarkers. These data demonstrate that coupling dynamic TIF proteomics with targeted serum proteomics in an animal model is a promising avenue for pursuing the discovery of tumor serum biomarkers.Biological significanceColorectal cancer (CRC) is one of the most dangerous diseases worldwide. However, few of CRC biomarkers possess satisfied specificity and sensitivity in clinical practices. Exploration of more CRC biomarkers, especially in serum, is an urgent and also a time-consuming campaign in the CRC study. Our study demonstrates that quantitatively evaluating the phase-dependent proteins in colonic tissue interstitial fluids from AOM–DSS mice is a feasible and effective way for exploration of the CRC-related proteins and the potential serum biomarkers. We identified two proteins, LRG1 and TUBB5, which may be practicable in human clinical samples as CRC serum biomarkers. To sum up, this study provides a novel angle to explore the critical factors in tumorigenesis and a new pipeline for potential serum biomarker discovery and verification.