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目的:蛋白质组学分析被认为是一种寻找潜在生物标志物,尤其是肿瘤标志物的有效方法。二维电泳蛋白质组学分析是一种流行和行之有效的技术。使用二维微分凝胶电泳(2-D DIGE)的研究是为了解癌症病人和对照组不同的蛋白质表达,并从蛋白质在二维图像上分布模式发现潜在的肿瘤标志物。方法:选取5例胆管癌病人和5例对照组胆管标本作为实验样本。然后提取DIGE裂解的蛋白质。随后提取的蛋白质以3种不同CyDyes进行标记,并通过2-D DIGE分离。结果:2-D DIGE从肿瘤和对照组标本中分离出多种蛋白质,点状分布在长13 cm、pH值在3.0～10.0的固相pH梯度胶条上。经过DeCyder软件的生物变异分析模块分析,所有胆管癌病人有20处蛋白质点被发现显着升高,5处蛋白质点下调(配对t检验,P<0.05)。然后由MALDI-TOF在16处蛋白质点上鉴定出13种蛋白质。结论:本研究采用2-D DIGE的方法对照研究胆管癌蛋白质组学,取得了胆管癌的二维蛋白分布图。经质谱鉴定的13种差异表达蛋白可作为潜在的肿瘤标志物以备进一步研究。
Purpose: Proteomics analysis is considered as an effective method to find potential biomarkers, especially tumor markers. Two-dimensional electrophoresis proteomics analysis is a popular and proven technology. Two-dimensional differential gel electrophoresis (2-D DIGE) was used to understand the differential expression of proteins in cancer patients and controls, and to identify potential tumor markers from patterns of distribution of proteins on two-dimensional images. Methods: Bile duct specimens from 5 cholangiocarcinoma patients and 5 control subjects were selected as experimental samples. The DIGE-cleaved protein is then extracted. Subsequent extracted proteins were labeled with 3 different CyDyes and separated by 2-D DIGE. Results: 2-D DIGE isolated a large number of proteins from the tumor and control samples. The spots were distributed on a 13 cm long, pH-sensitive strip of pH-gradient 3.0 to 10.0. After analysis by DeCyder software, the bio-variation analysis module showed that 20 of the 93 cholangiocarcinoma patients were found to have significantly increased protein spots and 5 of the protein spots were down-regulated (paired t-test, P <0.05). Thirteen proteins were then identified by MALDI-TOF at 16 protein spots. Conclusion: In this study, 2-D DIGE was used to compare the proteomics of cholangiocarcinoma and to obtain a two-dimensional protein profile of cholangiocarcinoma. The 13 differentially expressed proteins identified by mass spectrometry serve as potential tumor markers for further study.