人胆管癌与正常人胆汁蛋白质组学研究

Proteomic analysis of cholangiocarcinoma bile and normal human bile

背景与目的:胆管癌是目前常见的消化道恶性肿瘤,由于大多数患者确诊时疾病已属中晚期,总体5年生存率极低,所以改善预后的关键在于早期诊断。近年来,蛋白质组学的迅猛发展,为检测肿瘤标志物提供了一种新技术。本研究采用蛋白质组学的相关技术,建立胆汁蛋白质组双向电泳图谱,筛选胆管癌患者胆汁和正常人胆汁中的差异表达蛋白,以发现可能用于早期诊断胆管癌的肿瘤标志物。方法:收集胆管癌患者和正常人胆汁标本,经脱盐、脱脂纯化处理后提取蛋白,进行二维电泳,应用PDQuest 8.02D图像软件对银离子染色的双向电泳图像进行分析,并对差异表达的蛋白质行质谱鉴定。结果:成功建立胆管癌和正常人胆汁的双向凝胶电泳图谱,其图谱上平均蛋白质点数分别为(352±23)个和(317±26)个。选取其中表达差异超过2倍的蛋白质斑点10个,行质谱分析和数据库检索,鉴定出5种有意义的蛋白质,包括(angiopoienrelated protein 1)、(fibroblast growth factor substrate 2)、(catenin,beta 1)、(kertain,typeⅠ cytoskeletal 16)和(ras-related protein 37)。从功能上分析这些差异蛋白质与癌细胞的发生、增殖、分化、转移等相关。结论:蛋白质组学能很好地显示胆管癌患者与正常人胆汁间的差异表达蛋白,研究鉴定的5种差异表达蛋白质可能为研究胆管癌的生物学行为提供新的分子标志物。

Background and purpose： At present, cholangiocarcinoma is one of the common digestive tract tumors worldwide. Most of the patients are diagnosed at advanced stage, and its 5-year survival rate is very poor.Early diagnosis may improve the prognosis of patients with cholangiocarcinoma. In recent years, with the development of proteomics, new technology provides detecting tumor markers. This study applied the cross-correlation technique of proteomics to establish a 2-DE human biliary map for the purpose of comparing differential expressing proteins from cholangiocarcinoma and normal bile, which we can identify as biomarkers for early diagnosis of cholangiocarcinoma. Methods： Both cholangiocarcinoma bile and normal bile were collected,then removed the bile salt and lipid ,extracted the protein ,and then did 2-DE electrophoresis. The 2-DE maps were visualized after silver staining and analyzed by PDQuest 8.0 2D software,and the differential expression proteins were identified by mass spectrometry. Results： Dimensional protein maps of either cholangiocarcinoma or matched normal bile were gained successfully.Gel-analysis software identified an average of 352±23 proteins in cholangiocarcinoma while 317±26 proteins in normal bile and statistical filtering yielded 10 spots of a 2-fold change,5 of which were identified by using mass spectrometry, including （angiopoien-related protein 1）,（fibroblast growth factor substrate 2）,（catenin, beta 1）,（kertain,type Ⅰ cytoskeletal 16）,（ras-related protein 37）. Functional analysis revealed that these proteins were associated with cancer cellular oncogenesis, proliferation,differentiation and metastasis. Conclusion： Proteomic analysis can identify the proteins variance in cholangiocarcinoma bile versus normal human bile as well as providing probable new biomarkers correlated with biological behavior of cholangiocarcinoma.