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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 ,比较蛋白质组学是一种筛选癌变相关分子标志物的可靠方法之
To screen molecular markers of the progression of squamous atypical hyperplasia in bronchial epithelium, two-dimensional electrophoresis (2 DE) was performed using modified deoxycholate trichloroaeticacid (DOC TCA) to purify the total protein of the bronchial epithelium. ImageMaster 2D analysis software, Student’s t test to identify differential protein spots, matrix assisted laser desorption / ionization time of flight mass spectrometry (MALDI TOF MS) to obtain corresponding peptide mass fingerprint (PMF), search database to identify differences Protein, thereby obtaining the 2 DE map of human atypical hyperplasia and invasive carcinoma of bronchial epithelium and the average protein spots of the gel (12 73 0 0 ± 4 3 31,132 6 0 0 ± 6 6 6 3) The average number of differentially expressed protein spots was 5600 ± 896. Thirty-eight differential protein spots were analyzed by PMF to identify some proteins related to cell growth, differentiation or tumorigenesis, followed by immunohistochemical detection of the differential proteins EGFR, c Jun, Mdm2 in two types of tissue expression, The results also showed similar expression differences in the differential expression of the protein in the process of atypical hyperplasia of bronchial epithelial.These proteins may participate in the carcinogenesis in different ways and the immunohistochemistry of EGFR, c Jun, Mdm2 The agreement between the results and the mass spectrometry results suggests that comparative proteomics is a reliable method of screening for cancer-associated molecular markers