胃癌血清蛋白标记物的筛选与鉴定  

Screen and identification of serum protein biomarkers in gastric cancer

作　　者：张辉[1] 李浩[1] 郭飞[1] 张大[1] 杨合英[1] 王家祥[1] 

机构地区：[1]郑州大学第一附属医院小儿外科,450052

出　　处：《中华胃肠外科杂志》2016年第3期317-322,共6页Chinese Journal of Gastrointestinal Surgery

基　　金：基金项目：国家自然科学基金（81172085）;河南省科技开放合作项目（132106000071）

摘　　要：目的筛选并鉴定胃癌患者血清中可能存在的特异性蛋白标记物，为胃癌的早期诊断提供更为完善的血清蛋白质指纹图谱模型。方法收集60例胃腺癌患者手术前后外周血标本。采用表面增强激光解吸电离飞行时间质谱（SELDI-TOF-MS）技术对胃癌患者（术前组、术后组、转移组）及正常对照组血清之间进行对比检测，筛选出胃癌患者血清中存在的特异性蛋白质或含量与正常人有差异的蛋白质。使用凝胶电泳（TRICINE．SDS．PAGE）技术对这些差异性蛋白质进行分离与纯化。通过基质辅助激光解吸电离串联飞行时间质谱（MALDI-TOF／TOF）技术对分离纯化的蛋白质进行鉴定，最终确定胃癌患者血清中具有特异性的蛋白标记物。结果胃癌术前组与正常组的质谱数据得到15个特异性m／z峰值（P〈0．01）；通过SVM算法筛选出Youden指数最高的组合模型，得到m／z峰位于6449．1的蛋白标记物；该蛋白胃癌术前组表达水平明显高于正常组（2299．3±2029．3比509．5±168.3，P〈0．01）。胃癌术后组与术前组的质谱数据经过处理和统计学分析后。得到P〈0．01的特异性m／z峰6个：参考Youden指数最高的组合模型，得到m／z峰位于6449．2的蛋白质标记物；该蛋白在胃癌术前组表达水平明显高于术后组（1247．9±685．0比476．5±157．8，P〈0．01）。胃癌术前组与转移组的质谱数据经过处理和统计学分析后，得到P〈0．01的特异性m／z峰12个；参考Youden指数最高的组合模型，得到蛋白质标记物m／z峰位于6448．9；该蛋白在胃癌转移组的表达强度明显高于术前组（1506．9±1036．5比649．7±621．0，P〈0．01）。通过MALDI．TOF／TOF平台对样本分离纯化及酶解后所得到的肽段混合物进行检测，m／z峰位于6449的蛋白质经过鉴定为载脂蛋白CⅢ（ApoCⅢ）。结论m／z峰位于6449的蛋白质经过鉴定为ApoCⅢ，考虑Objective To screen and identify the serum specific protein markers of patients with gastric cancer by proteomies technology, and to provide more comprehensive serum protein fingerprint model for the early diagnosis of gastric cancer. Methods Preoperative and postoperative blood samples were collected from 60 gastric cancer patients. Mass spectrometry （SELDI-TOF-MS） technology was used to detect and screen serum specific proteins in gastric cancer patients （preoperative group, postoperative group, metastasis group）, and the result was compared with normal control group. Gel electrophoresis （TRICINE SDS-OAGE） technology was applied in the separation and purification for those different protein. Matrix assisted laser desorption ionization tandem time-of-flight mass spectrometry （MALDI -TOF./TOF） technology was used in the identification for the proteins following separation andpurification. Result Mass spectrometry data of preoperative group and normal group resulted in 15 specific m/z peak （P 〈 0.01 ）. SVM screened by a combination of the highest index model Youden get m/z peak at 6 449.1 protein markers. The protein expression of preoperative group was significantly higher than that of normal group （2 299.3 ± 2 029.3 vs. 509.5 ±168.3, P〈 0.01）. Mass spectrometry data of preoperative group and postoperative group resulted in 6 specific m/z peak （P 〈 0.01）. SVM screened by a combination of the highest Youden index model indentified get m/z peak at 6 449.2 protein markers. The protein expression of preoperative group was significantly higher than that of postoperative group （1 247.9±685.0 vs. 476.5 ±157.8, P 〈 0.01）. Mass spectrometry data of preoperative group and metastasis group resulted in 12 specific m/z peak （P 〈 0.01 ）. SVM screened by a combination of the highest Youden index model indentified get m/z peak at 6 448.9 protein markers. The protein expression of metastasis group was higher than that of preoperative group（ 1 506.9 ±1 036.5 vs. 649.7 ± 621.0�

关 键 词：胃肿瘤 蛋白质组学 肿瘤标记物 载脂蛋白CⅢ 

分 类 号：R735.2[医药卫生—肿瘤]