激光捕获显微切割纯化的人结肠癌组织的定量蛋白质组学研究

发布时间：2018-04-05 21:15

本文选题：结肠癌　切入点：生物标志物　出处：《基因组学与应用生物学》2017年08期

【摘要】：为分析结肠上皮癌变进程中的差异表达蛋白质,筛选结肠癌诊断候选标志物,先采用激光捕获显微切割技术(LCM)纯化人正常结肠组织和结肠癌组织,再采用同位素标记对蛋白质进行相对和绝对定量(i TRAQ),然后结合强阳离子柱分离和反相液质联用(2D-LC-MS/MS)鉴定差异表达蛋白质。试验共鉴定了1 042个非冗余蛋白,筛选出与人结肠癌变相关的差异蛋白162个,其中在结肠癌中表达上调的蛋白质68个,表达下调的蛋白质94个,功能分析表明,这些差异蛋白质涉及内质网蛋白质加工、糖酵解/糖原异生、黏着斑细胞连接、调节微丝组装和细胞外基质受体相互作用等分子功能。进一步采用Western blotting及免疫组织化学技术(IHC)验证了其中差异蛋白S100A9的表达,证实了定量蛋白质组学结果的可靠性。研究表明:差异表达蛋白质与结肠癌变相关,并有可能成为结肠癌诊断的候选分子标志物,为进一步深入研究这些蛋白质在结肠癌发生和发展进程中的分子机制,为结肠癌的发病机制研究提供新线索。
[Abstract]:In order to analyze the differentially expressed proteins in the carcinogenesis of colon epithelium and to screen the candidate markers for the diagnosis of colon cancer, the normal colon tissue and colon cancer tissue were purified by laser capture microdissection (LCM).Then the relative and absolute quantification of the protein was carried out by isotope labeling, and then the differentially expressed protein was identified by strong cationic column separation and reversed phase liquid / mass spectrometry (RLC-MS).A total of 1,042 nonredundant proteins were identified and 162 differentially expressed proteins were screened out, including 68 up-regulated proteins and 94 down-regulated proteins in colon cancer.These differential proteins are involved in the processing of endoplasmic reticulum proteins, glycolysis / glycogen heterogeneity, adhesion of plaque cells, regulation of microfilament assembly and interaction of extracellular matrix receptors, and so on.Furthermore, Western blotting and immunohistochemical technique were used to verify the expression of differential protein S100A9, which confirmed the reliability of quantitative proteomics results.Studies have shown that differentially expressed proteins are associated with colon cancer and may become candidate molecular markers for the diagnosis of colon cancer. In order to further study the molecular mechanism of these proteins in the carcinogenesis and development of colon cancer,It provides a new clue for the study of the pathogenesis of colon cancer.
【作者单位】：中南大学湘雅医院卫生部肿瘤蛋白质组学重点实验室抗癌药物国家与地方联合工程实验室结构生物学与药物设计湖南省工程实验室;南华大学药学与生物科学学院湖南省分子靶标新药研究协同创新中心;广西科技大学医学院;
【基金】：国家重点基础研究发展计划(973计划)(2011CB910704、2014CBA02004) 国家自然科学基金(81272971、81202129、81372516) 湖南省教育厅资助科研项目(16C1409)共同资助
【分类号】：R735.35

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