摘要
目的研究人大肠癌高、低分化组织及癌旁正常黏膜之间蛋白质组表达差异,寻找和鉴定癌组织与正常黏膜之间以及高、低分化癌组织之间差异表达蛋白。方法收集新鲜大肠癌标本;根据病理诊断结果将大肠癌标本分为高分化癌组、低分化癌组,以癌旁正常肠黏膜作为对照。提取组织总蛋白,进行双向凝胶电泳,比较得到高分化与低分化组织之间表达差异点。将这些差异蛋白点进行肽指纹图分析及网上数据库鉴定。应用Westernblot验证蛋白质组筛选结果。结果在两组癌标本中均存在钙网硬蛋白前体、O98007蛋白类似物的特异表达以及AAH10915、AAH75839蛋白表达上调;在高分化癌组中存在E980237蛋白的特异表达以及三磷酸异构酶、肝脂肪酸结合蛋白表达上调;在低分化癌组中存在2HHEB(氧化血红蛋白变异体)和锌指蛋白312类似物的特异表达以及Q9TQL5蛋白(Fragment)类似物表达上调。Westernblot检测HSP27和LFBP表达的结果与蛋白质组分析结果一致。结论高分化癌组织与低分化癌组织之间存在蛋白质组差异表达,这些蛋白与大肠癌细胞分化有关。
[Objective] The study aims to research the proteomie differences among the high and the low differentiation colorectal carcinoma tissues and the normal mucous epithelium by using two-dimensional gel electrophoresis （2-DE） and mass spectrometry. [Methods] In sample preparation, the fresh carcinoma tissue was cut off from the specimen immediately after the specimen was resected from the body. The tissues were classified into three groups according to the postoperative pathological diagnosis： high differentiation carcinoma, low differentiation carcinoma and normal epithelium. In the first part of our experiment, protein separating was performed using two-dimensional gel electrophoresis （2-DE） with Amersham 18cm linear pH 3-10 immobilized pH gradient （IPG） strips. After silver-staining, the images of the gels were acquired by the scanner and then analyzed to find the differentially expressed protein-spots in each group. In the second part of the experiment, the differentially expressed protein-spots was incised from the gels and digested by trypsin. We acquired the peptide mass fingerprintings（PMF） by matrix assisted laser desorption/ionization time-of-flight mass spectrometry（MALDI-TOF-MS） and identified the proteins by data searching in the Mascot-database. Protein expression were assayed by western blot. [Results] We detected calreticulin precursor and O98007-1ike protein expressed exclusively and AAH10915 和 AAH75839 increased in both the two cancer groups. In high differentiation group, E980237 and tubulin beta chain expressed exclusively and triosephosphate isomerase and fatty acid-binding protein increased. In low differentiation group, 2HHEB and Zinc finger protein312-1ike protein expressed exclusively and Q9TQLS-hke protein increased （over than 5 fold）. The HSP27 and LFBP expression by western blot were in accordance with results by proteomic methods. [ Conclusion ] Some differentially expressed proteins existed among high and the low differentiation colorectal carcinoma as well as
出处
《中国现代医学杂志》
CAS
CSCD
北大核心
2006年第13期2038-2040,共3页
China Journal of Modern Medicine
关键词
大肠癌
组织分化
蛋白质组学
colorectal carcinoma
differentiation
proteomics
