Uzozie Anuli Christiana, Selevsek Nathalie, Wahlander Asa, Nanni Paolo, Grossmann Jonas, Weber Achim, Buffoli Federico, Marra Giancarlo
From the ‡Institute of Molecular Cancer Research, University of Zurich, Switzerland.
§Functional Genomics Center Zurich, University/ETH Zurich, Zurich, Switzerland.
Mol Cell Proteomics. 2017 Mar;16(3):407-427. doi: 10.1074/mcp.M116.062273. Epub 2017 Jan 4.
Targeted proteomic methods can accelerate the verification of multiple tumor marker candidates in large series of patient samples. We utilized the targeted approach known as selected/multiple reaction monitoring (S/MRM) to verify potential protein markers of colorectal adenoma identified by our group in previous transcriptomic and quantitative shotgun proteomic studies of a large cohort of precancerous colorectal lesions. We developed SRM assays to reproducibly detect and quantify 25 (62.5%) of the 40 selected proteins in an independent series of precancerous and cancerous tissue samples (19 adenoma/normal mucosa pairs; 17 adenocarcinoma/normal mucosa pairs). Twenty-three proteins were significantly up-regulated ( = 17) or downregulated ( = 6) in adenomas and/or adenocarcinomas, as compared with normal mucosa (linear fold changes ≥ ±1.3, adjusted value <0.05). Most changes were observed in both tumor types (up-regulation of ANP32A, ANXA3, SORD, LDHA, LCN2, NCL, S100A11, SERPINB5, CDV3, OLFM4, and REG4; downregulation of ARF6 and PGM5), and a five-protein biomarker signature distinguished neoplastic tissue from normal mucosa with a maximum area under the receiver operating curve greater than 0.83. Other changes were specific for adenomas (PPA1 and PPA2 up-regulation; KCTD12 downregulation) or adenocarcinoma (ANP32B, G6PD, RCN1, and SET up-regulation; downregulated AKR1B1, APEX1, and PPA1). Some changes significantly correlated with a few patient- or tumor-related phenotypes. Twenty-two (96%) of the 23 proteins have a potential to be released from the tumors into the bloodstream, and their detectability in plasma has been previously reported. The proteins identified in this study expand the pool of biomarker candidates that can be used to develop a standardized precolonoscopy blood test for the early detection of colorectal tumors.
靶向蛋白质组学方法能够加速在大量患者样本中对多个肿瘤标志物候选物的验证。我们采用了名为选择/多反应监测（S/MRM）的靶向方法，以验证我们团队在先前对一大群结直肠癌前病变进行的转录组学和定量鸟枪法蛋白质组学研究中所鉴定出的结直肠腺瘤潜在蛋白质标志物。我们开发了SRM分析方法，以便在一系列独立的癌前和癌组织样本（19对腺瘤/正常黏膜；17对腺癌/正常黏膜）中可重复地检测和定量40种选定蛋白质中的25种（62.5%）。与正常黏膜相比，23种蛋白质在腺瘤和/或腺癌中显著上调（n = 17）或下调（n = 6）（线性倍数变化≥±1.3，校正P值<0.05）。大多数变化在两种肿瘤类型中均有观察到（ANP32A、ANXA3、SORD、LDHA、LCN2、NCL、S100A11、SERPINB5、CDV3、OLFM4和REG4上调；ARF6和PGM5下调），并且一个五蛋白生物标志物特征能够将肿瘤组织与正常黏膜区分开来，受试者工作特征曲线下的最大面积大于〇.83。其他变化则是腺瘤特异性的（PPA1和PPA2上调；KCTD12下调）或腺癌特异性的（ANP32B、G6PD、RCN1和SET上调；AKR1B1、APEX1和PPA1下调）。一些变化与一些患者或肿瘤相关表型显著相关。23种蛋白质中的22种（96%）有可能从肿瘤释放到血液中，并且先前已有关于它们在血浆中可检测性的报道。本研究中鉴定出的蛋白质扩大了可用于开发标准化结肠镜检查前血液检测以早期检测结直肠肿瘤的生物标志物候选物库。
