O'Neill J Robert, Pak Hui-Song, Pairo-Castineira Erola, Save Vicki, Paterson-Brown Simon, Nenutil Rudolf, Vojtěšek Bořivoj, Overton Ian, Scherl Alex, Hupp Ted R
From the ‡Edinburgh Cancer Research Centre at the Institute of Genetics and Molecular Medicine, Edinburgh University; Robert.o'
§Department of Surgery, Royal Infirmary of Edinburgh.
Mol Cell Proteomics. 2017 Jun;16(6):1138-1150. doi: 10.1074/mcp.M116.065078. Epub 2017 Mar 23.
Esophageal cancer is the eighth most common cancer worldwide and the majority of patients have systemic disease at presentation. Esophageal adenocarcinoma (OAC), the predominant subtype in western countries, is largely resistant to current chemotherapy regimens. Selective markers are needed to enhance clinical staging and to allow targeted therapies yet there are minimal proteomic data on this cancer type. After histological review, lysates from OAC and matched normal esophageal and gastric samples from seven patients were subjected to LC MS/MS after tandem mass tag labeling and OFFGEL fractionation. Patient matched samples of OAC, normal esophagus, normal stomach, lymph node metastases and uninvolved lymph nodes were used from an additional 115 patients for verification of expression by immunohistochemistry (IHC).Over six thousand proteins were identified and quantified across samples. Quantitative reproducibility was excellent between technical replicates and a moderate correlation was seen across samples with the same histology. The quantitative accuracy was verified across the dynamic range for seven proteins by immunohistochemistry (IHC) on the originating tissues. Multiple novel tumor-specific candidates are proposed and EPCAM was verified by IHC.This shotgun proteomic study of OAC used a comparative quantitative approach to reveal proteins highly expressed in specific tissue types. Novel tumor-specific proteins are proposed and EPCAM was demonstrated to be specifically overexpressed in primary tumors and lymph node metastases compared with surrounding normal tissues. This candidate and others proposed in this study could be developed as tumor-specific targets for novel clinical staging and therapeutic approaches.
食管癌是全球第八大常见癌症,大多数患者在确诊时已患有全身性疾病。食管腺癌(OAC)是西方国家的主要亚型,对当前的化疗方案大多具有抗性。需要选择性标志物来加强临床分期并实现靶向治疗,但关于这种癌症类型的蛋白质组学数据极少。经过组织学检查后,对来自7名患者的OAC以及匹配的正常食管和胃样本的裂解物进行串联质谱标签标记和离线凝胶分级分离后,再进行液相色谱串联质谱分析(LC MS/MS)。另外从115名患者中获取了与患者匹配的OAC、正常食管、正常胃、淋巴结转移灶和未受累淋巴结的样本,用于通过免疫组织化学(IHC)验证表达情况。在所有样本中鉴定并定量了六千多种蛋白质。技术重复之间的定量重现性极佳,相同组织学的样本之间存在中等程度的相关性。通过对原始组织进行免疫组织化学(IHC),在七种蛋白质的动态范围内验证了定量准确性。提出了多个新的肿瘤特异性候选物,并通过免疫组织化学(IHC)验证了上皮细胞黏附分子(EPCAM)。这项对OAC的鸟枪法蛋白质组学研究采用了比较定量方法,以揭示在特定组织类型中高表达的蛋白质。提出了新的肿瘤特异性蛋白质,并且证明与周围正常组织相比,EPCAM在原发性肿瘤和淋巴结转移灶中特异性过表达。本研究中提出的这一候选物及其他候选物可开发为用于新的临床分期和治疗方法的肿瘤特异性靶点。