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目的:筛选鼻咽癌(nasopharyngeal carcinoma,NPC)的差异表达蛋白质,为寻找NPC的分子标志物提供科学依据。方法:采用激光捕获显微切割技术(laser capture microdissection,LCM)分别从NPC组织和正常鼻咽上皮组织(normal nasopharyngeal epithelial tissue,NNET)中纯化NPC细胞和正常鼻咽上皮细胞(normal nasopharyngeal epithelial cells,NNEC),应用二维凝胶电泳(2-DE)分离LCM纯化细胞的蛋白质,图像分析识别差异表达的蛋白质点,基质辅助激光解吸电离飞行时间质谱(MALDI-TOF-MS)和电喷雾电离串联质谱(ESI-Q-TOF-MS)鉴定差异蛋白质点,Western印迹和组织芯片(tissue microarray,TMA)免疫组织化学验证差异蛋白鳞状细胞癌抗原1(SCCA1)在两种组织中的表达水平。结果:建立了LCM纯化的NPC细胞和NNEC的2-DE图谱,质谱鉴定了36个差异蛋白质,其中20个蛋白只在NPC表达或表达上调,16个蛋白在NPC中表达下调或缺失。Western印迹和组织芯片免疫组织化学结果显示:SCCA1在NPC中的表达水平较NNET明显下调,与比较蛋白质组学结果一致。结论:36个差异蛋白可能与NPC的发病有关,为筛选NPC分子标志物奠定了基础。
Objective: To screen differentially expressed proteins of nasopharyngeal carcinoma (NPC) and provide a scientific basis for finding molecular markers of NPC. Methods: NPC cells and normal nasopharyngeal epithelial cells (NPCs) were purified from NPC tissues and normal nasopharyngeal epithelial tissue (NNET) by laser capture microdissection (LCM) NNEC), proteins purified from LCM-purified cells were separated using 2-dimensional gel electrophoresis (2-DE), image analysis identified differentially expressed protein spots, matrix-assisted laser desorption ionization time of flight mass spectrometry (MALDI-TOF- MS), and electrospray ionization tandem The differentially expressed proteins were identified by ESI-Q-TOF-MS. Western blotting and tissue microarray (TMA) immunohistochemistry were used to confirm the expression of SCCA1 in both tissues. RESULTS: A 2-DE map of LCM-purified NPC cells and NNECs was established. Thirty-six differential proteins were identified by mass spectrometry. Twenty of these proteins were up-regulated or up-regulated only in NPC and down-regulated or down-regulated in NPC. Western blotting and tissue chip immunohistochemistry results showed that the expression level of SCCA1 in NPC was significantly lower than that in NNET, which was consistent with the comparative proteomics results. Conclusion: Thirty-six differential proteins may be related to the pathogenesis of NPC, which lays the foundation for the screening of NPC molecular markers.