结肠癌组织蛋白质组成分的双向凝胶电泳分析

PROTEOME ANALYSIS OF COLON CANCER BY TWO-DIMENSIONAL ELECTROPHORESIS

目的分析结肠癌病人癌组织、癌旁组织及正常结肠黏膜组织双向凝胶电泳的蛋白质表达图谱,寻找差异表达的蛋白质。方法采用固相pH梯度双向凝胶电泳分离结肠癌病人配对癌组织、癌旁组织及正常结肠黏膜组织的总蛋白,银染显色,PDQuest 2DE软件分析差异表达的蛋白质点。结果获得了重复性较好的双向凝胶电泳银染图谱。图像分析显示,3组胶的平均匹配率分别为:癌组织87.3%、癌旁组织80.3%、正常结肠黏膜组织84.0%。等电聚焦方向上的平均偏差为(1.16±0.29)mm,十二烷基硫酸钠-聚丙烯酰胺凝胶电泳(SDS-PAGE)方向上的平均偏差为(1.00±0.38)mm。差异分析共获得164个差异表达的蛋白质点。结论双向凝胶电泳分离结肠癌组织的总蛋白可获得重复性较好的结果。获得的差异蛋白质点为寻找结肠癌早期诊断的分子标记物提供了理论依据。

Objective To study the protein expression profiles of colon cancer mucosa, mucosa adjacent to cancer, and normal colon mucosa and detect the differential expression proteins. Methods Immobilized pH gradient two dimensional polyacrylamide gel electrophoresis （2 DE）, silver staining and PDQuest 2 DE analysis software were used to separate and analyze the pro teome of cancer mucosa, mucosa adjacent to cancer and normal mucosa. Results The 2 DE images were analyzed by PDQuest 2 DE software. The mean matching rates of the cancer, paraneoplastic and normalmucosa were 87.3%,80.3% and 84.0%, respec tively. In IEFdirection, the average deviation was （1.16±0.29） mm, in SDS-PAGE direction, the deviation was （1.00±0.38） mm. Using subtractive analyses of protein patterns of the cancer, paraneoplastic and normal mucosa, 164 different expression pro rein spots were screened. Conclusion Good reproducibility and resolution could be obtained by applying immobilized pH gradient two-dimensional polyacrylamide gel electrophoresis to separate and analyze the proteome in the tissues mentioned above. Proteins screened provide an evidence for searching markers of early detection of colon cancer.