早期肝癌甲胎蛋白阴性患者血清标志物的筛选和验证  

Screening and Identification of Serum Markers forα-Fetoprotein-Negative Patients with Early Stage Hepatocellular Carcinoma

作　　者：舒宏[1,2] 曹昭[3] 张舒[2] 甘淋[2] 秦雪[3] 刘银坤[2] 

机构地区：[1]广西医科大学附属肿瘤医院检验科,南宁530021 [2]复旦大学肝癌研究所,上海200032 [3]广西医科大学第一附属医院检验科,南宁530021

出　　处：《中国临床医学》2015年第4期451-455,共5页Chinese Journal of Clinical Medicine

基　　金：国家传染病重大专项基金资助项目(编号:2012ZX10002012-002)

摘　　要：目的:筛查甲胎蛋白(α-fetoprotein,AFP)阴性的肝癌患者和肝硬化患者血清中的差异表达蛋白。方法:选择6例AFP阴性的肝癌患者和6例AFP阴性的肝硬化患者。应用MARS系统分离获得各组血清标本中的低丰度蛋白,采用双向荧光差异凝胶电泳(two-dimensional fluorescence difference gel electrophoresis,2D-DIGE)技术筛选两组间有表达差异的蛋白,并用蛋白质印迹法加以验证。结果:2D-DIGE技术共鉴定出29种差异蛋白,其中AFP阴性肝癌患者血清中高表达的蛋白有22种,低表达的蛋白有7种。差异蛋白包括激肽原1、四连接素、基膜聚糖蛋白、补体1r和锌α2糖蛋白等。蛋白质印迹法证实,AFP阴性的肝癌患者血清中锌α2糖蛋白表达水平明显低于AFP阴性的肝硬化患者(P<0.05),同DIGE结果一致。结论:采用蛋白质组学技术筛查出的一系列在AFP阴性肝硬化和AFP阴性肝癌患者血清中有表达差异的蛋白有望作为AFP阴性肝癌患者早期诊断和辅助诊断的分子标志物。Objective：To screen and identify the differentially expressed proteins in the serum of α-fetoprotein （AFP）-negative patients with liver cancer and AFP-negative patients with liver cirrhosis. Methods： Six patients with AFP-negative liver cancer and 6 patients with AFP-negative liver cirrhosis were enrolled. The low abundance proteins in serum samples from each group were isolated by MARS system. Then the differentially expressed proteins between the two groups were screened by two-di- mensional fluorescence difference gel electrophoresis （2D-DIGE）technology. Furthermore, Western blotting was used to verify the differentially expressed proteins. Results： Totally, 29 differentially expressed proteins have been identified by 2D-DIGE technology. Among those proteins, 22 proteins were over-expressed in the serum from AFP-negative liver cancer patients, while 7 proteins were down-expressed. These differential proteins included kininogen 1, tetranectin, lumican, complement 1R and zinc-alpha-2-glycoprotein （ZAGP）. By Western blotting, it was confirmed that the expression of ZAGP in patients with AFP-negative liver cancer was significantly lower than that in patients with AFP-negative liver cirrhosis （P〈0. 05）, which was in accordance with the result of DIGE. Conclusions： A series of differentially expressed proteins were identified in the serum from AFP-negative patients with liver cirrhosis or liver cancer by using proteomic technology. These proteins might be used as molecular markers for the early diagnosis and assistant diagnosis of AFP-negative patients with liver cancer.

关 键 词：肝肿瘤 肝硬化 双向荧光差异凝胶电泳 蛋白质印迹法 甲胎蛋白 

分 类 号：R735.7[医药卫生—肿瘤]