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背景与目的:肺鳞癌的发生发展是一个多因素、多步骤的过程,虽然从基因和转录水平已对肺癌进行了许多成功的研究,但其癌变机制仍不十分明确,目前尚缺乏有效的用于肺鳞癌早期诊断和预后监测的特异性分子标志物。本研究的目的是利用蛋白质组学方法,建立分辨率高和重复性好的人肺鳞癌组织及其癌旁正常支气管上皮组织的双向凝胶电泳图谱,并识别鉴定其差异表达的蛋白质。方法:利用固相pH梯度(immobilizedpHgradient,IPG)双向凝胶电泳(two-dimensionalelectrophoresis,2-DE),分离人肺鳞癌组织及癌旁正常支气管上皮组织的总蛋白质,用图像分析软件比较分析,以识别差异表达的蛋白质;应用质谱仪(massspectrometry)得到相应的肽质指纹图谱(peptidemassfingerprint,PMF),然后搜索数据库鉴定部分差异蛋白质点。结果:(1)建立双向凝胶电泳图谱:癌组织和癌旁正常支气管上皮组织3块凝胶的平均蛋白质点数分别为1349±67和1297±73,平均匹配的点数分别为1235±48和1183±56,匹配率达91.5%和91.2%;同一癌组织的3块胶在蛋白质点位置上有较好的重复性,不同胶间蛋白质点在等电聚焦(Isoelectricfocusing,IEF)方向的偏差是(0.873±0.125)mm,在十二烷基磺酸钠聚丙烯酰胺凝胶电泳SDS-PAGE方向上的偏差为(1.025±0.213)mm。(2)肽质指?
BACKGROUND & OBJECTIVE: The occurrence and development of squamous cell carcinoma of the lung are a multifactorial and multi-step process. Although many successful studies have been done on lung cancer at the gene and transcription levels, the mechanism of carcinogenesis remains unclear. At present, there is a lack of effective Specific molecular markers for the early diagnosis and prognosis monitoring of squamous cell lung cancer. The purpose of this study was to establish a two-dimensional gel electrophoresis pattern of human lung squamous cell carcinoma and its adjacent normal bronchial epithelium with high resolution and good reproducibility by using proteomics methods and to identify and identify differentially expressed proteins. Methods: The total proteins of human lung squamous cell carcinoma and adjacent normal bronchial epithelial tissues were isolated by immobilized pH gradient (IPG) two-dimensional electrophoresis (2-DE), and compared with image analysis software. To identify differentially expressed proteins; mass spectrometry (mass spectrometry) was used to obtain the corresponding peptide mass fingerprint (PMF), and then the database was searched for partial differential protein spots. Results: (1) The establishment of two-dimensional gel electrophoresis: the average protein spots of three gels in cancer tissues and adjacent normal bronchial epithelial tissues were 1349 ± 67 and 1297 ± 73, respectively, and the average matching points were 1235 ± 48 and 1183 ± 56, and the match rates were 91.5% and 91.2% respectively. The three gums of the same cancer tissue had good repeatability at the protein spots, and the deviation of the protein spots in different gums in the direction of isoelectric focusing (IEF) was ( 0.873 ± 0.125) mm, and the deviation of sodium dodecyl sulfate polyacrylamide gel electrophoresis SDS-PAGE was (1.025 ± 0.213) mm. (2) Peptide means?