CAJ | 학술논문

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1 r 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。
목적：：사사갑태단백(α-fetoprotein,AFP)음성적간암환자화간경화환자혈청중적차이표체단백。방법：선택6례 AFP음성적간암환자화6례 AFP 음성적간경화환자。응용 MARS 계통분리획득각조혈청표본중적저봉도단백,채용쌍향형광차이응효전영(two-dimensional fluorescence difference gel electrophoresis,2D-DIGE)기술사선량조간유표체차이적단백,병용단백질인적법가이험증。결과：2D-DIGE 기술공감정출29충차이단백,기중 AFP 음성간암환자혈청중고표체적단백유22충,저표체적단백유7충。차이단백포괄격태원1、사련접소、기막취당단백、보체1 r 화자α2당단백등。단백질인적법증실,AFP음성적간암환자혈청중자α2당단백표체수평명현저우 AFP 음성적간경화환자(P <0.05),동 DIGE 결과일치。결론：채용단백질조학기술사사출적일계렬재 AFP 음성간경화화 AFP 음성간암환자혈청중유표체차이적단백유망작위 AFP 음성간암환자조기진단화보조진단적분자표지물。
Objective:To screen and identify the differentially expressed proteins in the serum ofα-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 1 R 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.