激光捕获显微切割技术结合iTRAQ标记定量蛋白质组在结肠癌筛查中的应用研究

The application of LCM combined with iTRAQ quantitative proteomic technology in colon cancer

目的采用激光捕获显微切割技术(LCM)结合iTRAQ标记定量蛋白质组技术,筛查结肠癌间质的差异表达蛋白质。方法 (1)选择12对结肠腺癌标本为观察组,其配对正常结肠黏膜(与结肠癌标本至少相距10 cm以上)为对照组,采用LCM技术分别纯化其间质作为生物样本;(2)iTRAQ标记定量蛋白质组技术鉴定两组间的差异表达蛋白质;(3)采用Geneontology生物信息学软件(http://pantherdb.org/)对差异表达蛋白质进行细胞成分、生物学途径和分子功能分析。结果 (1)鉴定了70个差异表达蛋白,其中37个蛋白在结肠癌间质中高表达,33个蛋白在结肠癌间质中表达降低;(2)从生物学途径、细胞成分、分子功能三个方面对差异表达蛋白质进行分析。结论 (1)首次采用iTRAQ标记定量蛋白质组学和LCM结合的方法筛查结肠癌间质差异表达蛋白质;(2)GO功能分析显示了差异表达蛋白质的细胞成分、分子功能以及生物学功能。

Objective To search differentially expressed proteins in colon cancer stroma （CS） by laser capture microdis- section （LCM） combined with iTRAQ marker quantitative proteomic technique. Methods ①12 CS and adenocarcinoma tissues were selected as observation group, and normal mucosa （〉10 cm away from adenocarcinoma） was selected as control group, the tissues were separated by LCM. ②iTRAQ quantitative proteomic technology was used to identified differentially expressed proteins （DEPs）. ③ GENEONTOLOGY functional annotation （GO） was used for bioinformatics analysis （Cellular components, biological pathways and molecular functional analysis）. Results ①70 DEPs were identi- fied in CS （37 upregulated and 33 downregulated）. ②The differentially expressed proteins were analyzed from three as- pects： biological pathway, cell composition and molecular functio. Conclusion ① For the first time, 70 DEPs in CS by LCM and iTRAQ quantitative proteomic technology are identified. ② GO can be used to analysis the biology function of the 70 DEPs and provided important values to the pathology of colon cancer.