胃癌组织及癌旁组织蛋白质双向电泳图谱的差异分析

Differential Analysis of Two-dimensional Gel Electrophoresis Profiles of Gastric Carcinoma and Paired Normal Gastric Mucosa

目的 利用蛋白质组学方法建立分辨率高和重复性好的人胃癌组织及其正常胃黏膜组织的双向凝胶电泳图谱 ,分析其差异表达的蛋白质 ,为进一步寻找胃癌标志物奠定基础。方法 采用固相pH梯度双向凝胶电泳分离人胃癌组织及正常胃黏膜组织的总蛋白质 ,凝胶经银染显色后 ,ImagingMaster 2D图像分析软件进行比较分析 ,识别差异表达的蛋白质。结果 ⑴癌组织和正常胃黏膜组织凝胶的平均蛋白质点数分别为 10 49± 67和 10 97± 73 ,平均匹配的点数分别为 83 5± 48和 95 3± 5 6,匹配率分别为79 6%和 86 7%。⑵通过比较分析 5例胃癌组织及正常胃黏膜组织的双向凝胶电泳图谱 ,得到平均匹配蛋白点数为 769± 45 ,差异表达蛋白点数为 81个 ,其中 17个点仅在癌组织中表达 ,2 4个点仅在正常胃黏膜中表达 ,2 6个点在癌组织中为高表达 ,14个点在癌组织中为低表达。结论 本研究建立了分辨率高且重复性较好的人胃癌组织与正常胃黏膜组织的双向凝胶电泳图谱 ,发现两者间存在一些差异表达的蛋白质 ,为进一步筛选胃腺癌特异性的分子标志物打下了坚实的基础。

Objective To establish two-dimensional electrophoresis profiles with high resolution and reproducibility from human gastric carcinoma and paired normal gastric mucosa tissues and to analyse the differential expression proteins between the two types of the tissues. Methods The total proteins of gastric carcinoma and paired normal gastric mucosa tissues were seperated by immobilized pH gradient-based two-dimensional gel electrophoresis (2-DE).After silver staining, the differential expression proteins were analyzed by using Imaging Master 2D analysis software. Results ⑴Average protein spots were 1049±67, 1097±73 in gastric carcinoma and paired normal gastric mucosa respectively,the matched spots were 835±48, 953±56 in both the two types of tissues respectively, and the average matching rate was 79.6%, 86.7% respectively. ⑵A total of 769±45 protein spots were matched between the electrophoretic maps of 5 gastric carcinoma and paired normal gastric mucosa tissues. The 81 differential protein spots were identified between the two types of the tissues. 17 protein spots were expressed only in the gastric carcinoma and 24 protein spots were expressed only in the normal gastric mucosa. The expression levels of 26 protein spots in the gastric carcinoma were higher than those in the normal gastric mucosa, and the expression levels of 14 protein spots in the gastric carcinoma were lower than those in the normal gastric mucosa. Conclusion In this study, the well-resolved,reproducible 2-DE patterns of gastric carcinoma and paired normal gastric mucosa tissues were established and some differential proteins between the two types of tissues were found. These data will be helpful to further screen specific biomarkers of gastric carcinoma.