人支气管上皮不典型增生与浸润癌组织的比较蛋白质组学研究  

Comparative Proteomics Analysis of Dysplasia and Invasive Cancer Tissues in Human Bronchial Epithelia

作　　者：吴晓英[1,2,3] 陈主初[1,2] 肖志强[1] 李萃[1,2] 汪春年[4] 李建玲[1] 冯雪萍[1] 张鹏飞[1] 

机构地区：[1]中南大学湘雅医院卫生部肿瘤蛋白质组学重点实验室 [2]中南大学湘雅医学院肿瘤研究所,长沙410078 [3]中南大学基础医学院超微结构教研室,长沙410078 [4]中南大学基础医学院超微结构教研室

出　　处：《生物化学与生物物理进展》2004年第11期1006-1017,共12页Progress In Biochemistry and Biophysics

基　　金：国家重点基础研究发展规划项目 ( 973 ) ( 2 0 0 1CB5 10 2 0 7) ;教育部跨世纪优秀人才基金项目 (教育部科技函 [2 0 0 2 ] 48号 ) ;国家自然科学基金项目 ( 3 0 0 0 0 0 2 8;3 0 2 40 0 5 6;3 0 3 70 64 2 ) ;湖南省科技厅重大和重点科研项目 ( 0 4XK10 0 1;0 2SSS

摘　　要：为筛选支气管上皮鳞状不典型增生进展的分子标志物 ,采用改良的脱氧胆酸 三氯醋酸 (deoxycholate trichloroaeticacid ,DOC TCA)法提纯支气管上皮总蛋白质进行双向电泳 (two dimensionalelectrophoresis,2 DE) ,应用ImageMaster 2D分析软件、Student’st 检验识别差异蛋白质点 ,基质辅助激光解吸电离飞行时间质谱(matrix assistedlaserdesorption/ionizationtime of flightmassspectrometry ,MALDI TOF MS)得到相应的肽质指纹图 (peptidemassfingerprint,PMF) ,搜索数据库鉴定差异蛋白质 .由此获得人支气管上皮不典型增生和浸润癌组织的 2 DE图谱及其凝胶的平均蛋白质点数 (12 73 0 0± 4 3 31,132 6 0 0± 6 6 6 3) ,且两阶段间平均差异蛋白质点数为 5 6 0 0± 8 96 .取 38个差异蛋白质点进行PMF分析 ,鉴定出一些与细胞生长、分化或肿瘤发生等有关的蛋白质 ,随即应用免疫组化检测差异蛋白质EGFR、c Jun、Mdm2在两类组织中的表达 ,其结果也显示了类似的表达差异 .支气管上皮不典型增生恶性转化过程中存在蛋白质的差异表达 ,这些差异蛋白质可能以不同的方式参与了癌变过程 ,且EGFR、c Jun、Mdm2的免疫组化验证结果与质谱结果的一致性表明 。In order to examine molecular markers of dysplasia progression, improved deoxycholate-trichloroaetic acid DOC-TCA) extracted total protein of bronchial epithelial. After profiling squamous dysplasia versus invasive carcinoma of bronchial epithelium using two-dimensional electrophoresis, the patterns were compared and differential proteins were found with lmgeMaster software and Student. t-test ( P < 0.05). The selected differential protein-spots were identified by peptide mass fingerprint (PMF) based on matrix-assisted laser desorption ionization time-of -flight mass spectrometry (MALDI-TOF-MS) and database searching. The average spots for dysplasia and invasive carcinoma were ( 1 273.00 +/- 43.31) and ( 1 326. 00 +/- 66. 63), respectively. After matching, the number of spots of differentially expressed proteins between the dysplasia and invasive cancer tissues were (56. 00 +/- 8. 96). Thirty eight differential proteins were identified, some of which are known to be involved in regulating the processes of proliferation, differentiation and tumorigenesis. In particular, c-Jun was up-regulated in invasive cancer compared with dysplasia, and murine double minute gene 2 (Mdm2) was only expressed in invasive cancer. But truncated EGFR ( epidermal growth factor receptor) is reduced in invasive cancer. The subsequent immunchistochemical analysis of human tissues biopsies also showed a similar expression pattern. Therefore there are differentially expressed proteins during malignant transformation of bronchial epithelium from dysplasia to cancer. These differential proteins probably take part in carcinogenesis in different forms. The similar results of mass spectrometry and immunohistochemistry illustrated the use and reliability of comparative proteomics to screen relevant molecular markers of carcinogenesis.

关 键 词：比较蛋白质组学 人支气管上皮 差异表达蛋白质 免疫组化 分子标志物 

分 类 号：Q50[生物学—生物化学] Q51