卵巢癌组织的差异蛋白质研究

To Explore the Differentia Expression Protein of Ovarian Cancer with 2DE-MS

目的分析卵巢癌患者癌组织、癌旁组织及正常卵巢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卵巢癌患者配对癌组织、癌旁组织及正常卵巢黏膜组织的总蛋白,银染显色,PDQuest2DE软件分析差异表达的蛋白质点;凝胶经银染显色后,Bio-Rad凝胶扫描仪扫描,Imagemaster图像软件分析,差异蛋白质点用基质辅助激光解析电离-两级飞行时间串联质谱法进行鉴定。结果获得了重复性较好的双向凝胶电泳银染图谱。3组胶的平均匹配率分别为:癌组织83.4%、癌旁组织81.2%、正常卵巢黏膜组织88.1%。斑点位置偏差等电聚焦方向上为(1.21±0.27)mm,SDS-PAGE方向上为(1.03±0.28)mm。差异分析共获得164个差异表达的蛋白质点;有差异性表达的8种蛋白质中有4种为结合珠蛋白的亚型。结论双向凝胶电泳-质谱技术能分离卵巢癌组织的总蛋白,并获得重复性较好的结果;结合珠蛋白能为诊断卵巢癌提供诊断手段。

Objective To study the protein expression profiles of cancer mucosa, mucosa adjacent to cancer, and normal ovarian mucosa and detect the differential expression proteins. Methods Immobilized pH gradient two-dimensional polyacrylamide gel electrophoresis （2 DE）, silver staining and PD Quest 2 DE analysis software were used to separate and analyze the proteome of cancer mucosa, mucosa adjacent to cancer and normal mucosa. Results The 2DE images were analyzed by PD Quest 2DE software. The mean matching rate of the cancer, paraneoplastic and normal mucosa were 83.4%,81. 2% and 88.1%, respectively. In IEF direction, the average deviation was（1.21±0.27） mm, in SDS-PAGE direction, the deviation was （1.03±0.28）mm. Using subtractive analyses of protein patterns of the cancer, paraneoplastic and normal mucosa, 164 different expression protein spots were screened, 8 kinds of the significant differential proteins were selected and identified by MALDI-TOF-TOF, it was found that 4 kinds of them were hypotype of haptoglobin. Conehtsion Good reproducibility and resolution could be obtained by applying 2DE-MS to separate and analyze the proteome in the tissues mentioned above. Haptoglobin can provide the diagnosis methods for screening and provide the evidence for searching markers in early detection of ovarian cancer.