人肺鳞癌组织及癌旁组织蛋白质双向电泳图谱的差异分析

Differential Analysis of Two-Dimension Gel Electrophoresis Profiles of Human Lung Squamous Carcinoma and Tumor-Adjacent Tissue

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: Carcinogenesis of lung squamous carcinoma i s a complex process involving multiple events and steps. Though some molecular p athogenesis studies on human lung cancer have been undertaken successfully in ge ne (DNA) and transcription (mRNA) levels, the carcinogenic mechanism is still un clear. At present, there is no special molecular marker for early-stage diagnos is and prognosis evaluation. The objective of this study was to establish two-d imensional electrophoresis profiles with high resolution and reproducibility fro m human lung squamous carcinoma tissue and paired normal tumor-adjacent bronchi al epithelial tissue, and to identify differential expression proteins. METHODS: The total proteins of human lung squamous carcinoma tissue and paired normal tu mor-adjacent bronchial epithelial tissue were separated by immobilized pH gradi ent (IPG)-based two-dimensional gel electrophoresis (2-DE). The differential expression proteins were analyzed using image analysis software, then identified using mass spectrometry and database searching. RESULTS: (1) For tumor tissues, the average protein spots of 3 gels were 1349±67; and 1235±48 spots were matc hed with the average matching rate of 91.5%. For control, the average protein s pots of 3 gels were 1297±73; and 1183±56 spots were matched with the average m atching rate of 91.2%. The average position deviation of matched spots was 0.87 3±0.125 mm in IEF direction, and 1.025±0.213 mm in SDS-PAGE direction. (2) A total of 1069±45 spots were matched between the electrophoretic maps of 15 huma n lung squamous carcinoma tissue and paired normal tumor-adjacent bronchial epi thelial tissue. Forty differential proteins were identified by peptide mass fing erprint(PMF), some proteins were the products of oncogenes, and the others invol ve in the regulation of cell cycle and signal transduction. CONCLUSION: In this study, the well-resolved, reproducible 2-DE patterns of human lung squamous ca rcinoma and adjacent normal bronchial epithelial tissues were established and ce rtein differential proteins were characterized. These data will be helpful for s creening the biomarker to further study on human lung squamous carcinoma.