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目的:建立肺鳞癌(human lung squamous carcinoma,HLSC)组织与正常支气管上皮(normal humanbronchial epithelial,NHBE)组织的差异蛋白质表达谱。方法:运用激光捕获显微切割(laser capture microdissection,LCM)技术从HLSC及NHBE组织中分别纯化HLSC和NHBE细胞,应用同位素标记相对和绝对定量(isobaric tagsfor relative and absolute quantitation,iTRAQ)和二维液相色谱-串联质谱技术分离、鉴定HLSC和NHBE的差异表达蛋白质。结果:共鉴定了96个HLSC和NHBE之间的差异表达蛋白质,其中在HLSC中上调的蛋白49个、下调蛋白47个。采用Western印迹对其中的两个差异蛋白质HSPB1和CKB进行了表达水平验证,结果与定量蛋白质组学研究的结果一致。结论:鉴定的肺鳞癌差异表达蛋白质为研究HLSC癌变机制奠定了基础。
OBJECTIVE: To establish a differential protein expression profile of human lung squamous carcinoma (HLSC) tissues and normal human bronchial epithelial (NHBE) tissues. METHODS: HLSC and NHBE cells were purified from HLSC and NHBE tissues by laser capture microdissection (LCM). The isobaric tags for relative and absolute quantitation (iTRAQ) Separation and identification of differentially expressed proteins of HLSC and NHBE by phase chromatography-tandem mass spectrometry. RESULTS: A total of 96 differentially expressed proteins between HLSC and NHBE were identified, including 49 proteins up-regulated in HLSC and 47 proteins down-regulated. Two differential proteins, HSPB1 and CKB, were verified by Western blotting and the results were consistent with the results of quantitative proteomics studies. Conclusion: The identified differentially expressed proteins of lung squamous cell carcinoma lay the foundation for the study of carcinogenesis mechanism of HLSC.