激光捕获显微切割纯化的人正常支气管上皮与肺鳞癌组织的定量蛋白质组学研究

Quantitative proteomic study for purified human lung squamous carcinoma and normal human bronchial epithelial by laser capture microdissection

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 the differential protein profile for human lung squamous carcinoma （HLSC） and normal human bronchial epithelium （NHBE）. Methods Laser capture microdissection （LCM） was used to purify the target cells from HLSC tissues and matched NHBE. A stable isotope labeled strategy using iTRAQ together with 2 dimensional chromatography/hybrid quadrupole time of flight （2D-LC/Q-STAR） mass spectrometry was performed to separate and identify the differential expression proteins. Results A total of 96 differential expression proteins in the LCM-purified HLSC and NHBE were identified. Compared with NHBE, 49 proteins were upregulated while 47 proteins were downregulated in HLSC. The expression levels of the differential proteins HSPB1 and CKB were confirmed by Western blot analysis, which were consistent with the results of quantitative proteomics. Conclusion The different expression proteins in HLSC will provide useful information for exploring the carcinogenic mechanism of HLSC.