胃腺癌组织蛋白质双向电泳图谱分析

Differential Analysis of Two-dimensional Gel Electrophoresis Profiles for Gastric Adenocarcinoma

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腺癌组织及其正常胃黏膜组织的双向凝胶电泳图谱 ,并分析其差异表达的蛋白质 ,以期用于早期诊断。方法 采用固相pH梯度(immobilizedpHgradient,IPG)双向凝胶电泳 (two -dimensionalgelelectrophoresis ,2 -DE)分离人胃腺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 ( 1 )得到了分辨率较高、重复性较好的人胃腺癌组织和正常胃黏膜组织的双向凝胶电泳图谱 ,癌组织和正常胃黏膜组织凝胶的平均蛋白质点数分别为 1 0 4 9± 67和 1 0 97± 73,平均匹配的点数分别为 835± 48和 95 3± 5 6,匹配率分别为 79.6%和 86.7% ;同一组织凝胶在蛋白质点位置上有较好的重复性 ,不同凝胶间蛋白质点在等电聚焦 (isoelectricfocusing ,IEF)方向的偏差是 0 .873±0 .1 2 5mm ,在十二烷基磺酸钠聚丙烯酰胺凝胶电泳 (sodiumdodecylsulfate -polyacrylamidegelelectrophore sis ,SDS -PAGE)方向上的偏差为 1 .0 2 5± 0 .2 1 3mm。 ( 2 )通过比较分析 5例胃腺癌组织及正常胃黏膜组织的双向凝胶电泳图谱 ,得到平均匹配蛋白质点数为 769± 45 ,差异表达蛋白质点数为 81 ,其中 1

Objective To explore early-stage diagnostic method for gastric adenocarcinoma (GA), the objective of this investigation are the establishment of two-dimensional Gel electrophoresis profiles with high resolution and reproducibility from human GA and paired normal gastric mucosa tissues and the identification of differential expression proteins. Methods The total proteins of GA and paired normal gastric mucosa tissues were separated by means of immobilized pH gradient-based two-dimensional gel electrophoresis (2-DE).After AgNO 3 staining, the differential expression proteins were analyzed by using Imaging Master 2D analysis software. Results (1) The well-resolved,reproducible 2-DE patterns of GA and normal gastric mucosa were obtained. The results showed that average spots were 1 049±67, 1 097±73 in GA and normal gastric mucosa respectively,the matched spots were 835±48, 953±56 in them separately,the according average matching rate was 79.6%,86.7% respectively. The average position deviation of matched spots was 0.873±0.125 mm in IEF direction, and 1.025±0.213 mm in SDS-PAGE direction.(2) A total of 769±45 spots were matched between the electrophoretic maps of 5 GA and normal gastric mucosa tissues. The 81 differential protein spots were identified by Imaging Master 2D image analysis software between 5 cases of GA and paired control normal gastric mucosa. There were only 17 protein spots examined in GA and 24 protein spots examined in normal gastric mucosa. 26 protein spots in GA were higher expression than in normal gastric mucosa , 14 protein spots in GA were lower expression than in normal gastric mucosa. Conclusion In this study, the well-resolved,reproducible 2-DE patterns of GA and normal gastric mucosa tissues were established and some differential proteins were characterized. These data will be used to screen special biomarkers of GA for further.