应用蛋白质组学研究白斑差异表达蛋白  

Screening of Differential Expression Protein from Human Oral Leukoplakia and Normal Oral Mucosa Tissues by Two-dimensional Difference Gel Electrophoresis and Mass Spectrometry.

作　　者：王文梅[1] 郑春兰[1] 蒋文晖[1] 黄晓峰[1] 唐恩溢[1] 唐巍[1] 黄政[1] 段宁[1] 

机构地区：[1]南京大学医学院附属口腔医院口腔黏膜病科,江苏南京210008

出　　处：《口腔医学研究》2008年第3期255-258,共4页Journal of Oral Science Research

基　　金：江苏省社会发展科技项目(编号:BS2004504)

摘　　要：目的:通过筛选口腔白斑及正常黏膜组织的差异表达蛋白,为研究白斑发生机制提供实验依据。方法:收集口腔白斑组织及正常口腔黏膜组织,提取组织蛋白,进行二维电泳,选择在表达差异量较大的11个点进行质谱和生物信息学分析,确定所分析的蛋白质类型。结果:人口腔白斑及正常黏膜组织的二维电泳图谱的平均蛋白质点数分别为1726±67和1608±73,蛋白质点重复性较好。人口腔白斑与正常黏膜组织的双向凝胶电泳图谱,差异表达蛋白质点数为38个,其中16个点在白斑中为高标达,22个点在白斑中为低表达,选择11个表达差异量较大的蛋白质点进行质谱和生物信息学分析,鉴定出其中的5个点,分别为膜联蛋白A2,角蛋白8,角蛋白1,II型角蛋白亚基,免疫球蛋白(Ig)κ型轻链的恒定区(C区)片段等。结论:膜联蛋白A2,角蛋白8,角蛋白1,II型角蛋白亚基,Igκ型轻链的C区片段等在口腔白斑发生发展过程中发生了改变。Objective：To find early diagnostic biomarkers for oral leukoplakia by comparing differential expressing proteins from oral leukoplakia and normal oral mucosa tissues. Methods： Oral leukoplakia and paired normal oral mucosa tissues were collected, and tissue total protein was extracted. Differential proteome profiles were established and analysed by means of immobilized pH gradient-based two-dimesional polyacrylamide gel electrophoresis（ 2D-PAGE） and matrixassisted laser desorption/ionization time of flight mass spectrometry （MALDI-TOF-MS）. Results： The well-resolved, reproducible 2-DE patterns of oral leukoplakia and nomml oral mucosa were obtained. The results showed that average spots were 1726±67 and 1608±73 in oral leukoplakia and normal oral mucosa, respectively. The 38 differential protein spots were identified by Imaging Master 2D image analysis software between oral leukoplakia and paired control normal oral mucosa. There were 16 protein spots in oral leukoplakia which were higher expression than in normal oral mucosa, 22 protein spots in oral leukoplakia which were lower expression than in normal oral mucosa. 11 differential expressing proteins were analysed by mass spectrometry and bioinformation. 5 of them were well characterized including Annexin A2, cytokeratin 8, keratin 1, type II keratin subunit protein , and kappa chain c region （allotype Inv （1,2））-human （fragment）. Conclusion： Differential expression proteins can be candidate biomarkers for early diagnosis of oral leukoplakia.

关 键 词：口腔白 斑双向凝胶电泳 蛋白质组 膜联蛋白A2 角蛋白8 Ⅱ型角蛋白亚基 角蛋白1 Igκ型轻链的C区片段 

分 类 号：R78[医药卫生—口腔医学]