激光捕获显微切割技术纯化的人支气管上皮癌变各阶段组织的定量蛋白质组学研究

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为分析支气管上皮癌变进程中的差异表达蛋白质,筛选肺鳞癌早期诊断标志物,以人支气管上皮癌变各阶段组织为研究对象,先采用激光捕获显微切割技术(LCM)纯化人正常支气管上皮组织、鳞状化生、不典型增生、原位癌、浸润性肺鳞癌组织,再用同位素标记相对和绝对定量(iTRAQ)技术结合二维液相色谱串联质谱(2D LC-MS/MS)鉴定支气管上皮癌变进程中各阶段的差异表达蛋白质.结果共鉴定了1 036个蛋白质,筛选出102个与人支气管上皮癌变相关的差异蛋白质,在这些差异蛋白质中,有的在支气管上皮癌变过程中进行性上调,有的在支气管上皮癌变过程中进行性下调,有的呈阶段特异性改变.功能分析表明,这些差异蛋白质涉及代谢、细胞凋亡、增殖、分化、信号传导、转录、翻译、细胞黏附、免疫反应与发育等.Western blotting及免疫组织化学技术验证了其中2个差异蛋白(S100A9和CKB)的表达,证实了定量蛋白质组学结果的可靠性.研究结果提示:这些差异表达蛋白质与支气管上皮癌变相关,并可成为肺鳞癌的早期诊断标志物,进一步研究差异蛋白质的生物学功能,将有助于阐明支气管上皮的癌变机制,从而为肺鳞癌的早期诊断与发病机制研究提供新思路. To analyze the differentially expressed proteins in the process of bronchial epithelial carcinogenesis and to screen the early diagnostic markers of squamous cell carcinoma of the lung, the human bronchial epithelial carcinogenesis tissues at different stages were selected as the research object. Human normal bronchial epithelial tissue was purified by laser capture microdissection (LCM) , Squamous metaplasia, atypical hyperplasia, carcinoma in situ, and invasive lung squamous cell carcinoma tissues were identified by 2D LC-MS / MS with isotope labeled relative and absolute quantification (iTRAQ) A total of 10 036 proteins were identified and 102 differentially expressed proteins were detected in human bronchial epithelial carcinogenesis. Some of these differentially expressed proteins were involved in carcinogenesis of bronchial epithelial carcinogenesis Sexual up-regulation, some down-regulation in bronchial epithelial carcinogenesis, and some stage-specific changes.Functional analysis showed that these differential proteins involved in metabolism, apoptosis, proliferation, differentiation, signal transduction, transcription, translation, cell adhesion , Immune response and development, etc. Western blotting and immunohistochemistry confirmed that two of the differential proteins (S100A9 And CKB), confirming the reliability of quantitative proteomics results.The results suggest that these differentially expressed proteins are associated with bronchial epithelial carcinogenesis and may be an early diagnostic marker for squamous cell carcinoma of the lung, and further study the biology of differential proteins Function, will help clarify the mechanism of bronchial epithelial carcinogenesis, and thus provide a new idea for the early diagnosis and pathogenesis of squamous cell carcinoma of the lung.
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