高发区早期食管癌及癌前病变血清蛋白研究  

Serum proteomic study on early esophageal carcinoma and precancerous lesions and precancerous lesions in the high incidence area

作　　者：陈仙梅 袁丽[2] 郭硕[2] 于卫芳[3] 王士杰[2] 张立玮[2] 

机构地区：[1]宁夏人民医院消化科,银川750021 [2]河北医科大学第四医院内镜科 [3]河北医科大学第一医院内镜中心

出　　处：《中华临床医师杂志（电子版）》2014年第19期1-5,共5页Chinese Journal of Clinicians(Electronic Edition)

基　　金：河北省自然科学基金(C2010000638);河北省校科技合作课题(2011054278)

摘　　要：目的应用质谱技术构建高发区食管癌及癌前病变血清蛋白质指纹图谱模型,为食管癌早期诊断寻找生物指标,并探讨差异蛋白在食管癌发生发展中的作用及意义。方法收集食管癌、癌前病变血清样本共254例,根据病理分为低级别上皮内瘤变(LGIN)、高级别上皮内瘤变(HGIN)、进展期食管鳞癌(AEC)、健康对照组。采用弱阳离子交换纳米磁珠(WCX)对血清蛋白进行纯化,MALDI-TOF-MS质谱进行检测,构建诊断模型并进行盲法验证。结果三组病变组与对照组以及各组间比较有统计学差异蛋白峰146个,由14个蛋白峰建立食管癌及癌前病变血清蛋白指纹图谱模型,蛋白峰分别是32721、3403、1772、1071、1414、33312、32023、31043、4662、2245、1307、10013、6868、1234。对早期食管癌及前病变的特异性为90.91%(30/33),对LGIN的灵敏度为100%(31/31),HGIN灵敏度为89.66%(52/58),AEC的灵敏度为81.26%(26/32),盲法预测验证结果表明;该模型对食管癌前病变及癌的特异性为95.652%(22/23),对LGIN的敏感性为95.455%(21/22),HGIN敏感性为94.118%(32/34),AEC的敏感性为95.238%(20/21)。经ExPasy数据库检索蛋白峰为32023,1234,1307,推测为ZAG(zinc alpha2 glycoprotein),Glycosyltransferase-like protein LARGE2,Fibrinogen alpha chain。结论采用MALDI联合WCX建立诊断模型具有高的灵敏度及特异性。根据各组间差异蛋白变化,推测食管癌的发生可能起始于LGIN或更早,提示对于表达与肿瘤相关差异蛋白峰的个体应给予定期随访。Objective This study was to screen protein biomarkers in serum of esophageal carcinoma and precancerous lesions subjects using Mass Spectrometry in high incidence area of China. Boosting decision tree was constructed to identify normal from EC and precancerous lesions, to screen significant biomarkers for early diagnosis and explore the function in progression of esophageal cancer. Methods We recruited 254 subjects, Including low-grade intraepithelial neoplasia （LGIN）, high-grade intraepithelial neoplasia （HGIN）, advanced esophageal carcinoma （AEC） and normal control （NOR）. All serum samples were pretreated with weak cation exchange （WCX） magnetic beads and tested by MALDI-TOF-MS, finally construction of boosting decision tree and blinded testing. Results There were 146 protein peaks were detected between the three parents groups compared with normal, the decision tree classification were constructed by 14 protein peaks, which contain 32721, 3403, 1772, 1071, 1414, 33312, 32023, 31043, 4662, 2245, 1307, 10013, 6868, 1234. This decision algorithm correctly classified 90.91%（30/33） of normal control, 100% （31/31） of LGIN, 89.66% （52/58） of HGIN, and 81.26% （26/32） of AEC.In the blinded testing, the decision algorithm correctly classified 95.652%（22/23） of normal control, 95.455%（21/22） of LGIN, 94.118%（32/34） of HGIN, and 95.238%（20/21） of AEC. The peaks of 32023, 1234 and 1307 were speculated zinc alpha2 glycoprotein, glycosyltransferase like protein LARGE2 fibrinogen alpha chain and fibrinogen alpha chain by ExPasy database retrieval, others were unknown. Conclusion The diagnostic pattern are established by differently protein peaks could accurately recognize the patients groups and normal controls application of MALDI-TOF MS with high sensitivity and specificity. We speculate that the occurrence of esophageal carcinoma began in the LGIN or much earlier, the patients that have cancer-related proteins in serum should be follow up.

关 键 词：食管肿瘤 癌前病变 蛋白质组学 基质辅助激光解吸电离飞行时间质谱 

分 类 号：R735.1[医药卫生—肿瘤]