联合应用激光捕获显微切割技术及乙酰化稳定同位素标记定量技术在结直肠癌蛋白质组学研究中的价值
Role of Quantitative Proteomic Strategy by Laser Capture Microdissection Combined with Acetylation Stable Isotope Labeling in Colorectal Cancer
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摘要: 目的 应用蛋白质组学技术,建立结直肠癌蛋白质组差异表达谱,筛选结直肠癌发生发展相关差异蛋白质。方法应用激光捕获显微切割技术,获取20例结直肠腺癌及配对的正常黏膜、20例绒毛状腺瘤、8例结直肠癌肝转移灶的靶细胞;然后应用基于乙酰化稳定同位素标记的定量蛋白质组学策略寻找差异蛋白。结果获得具有定量信息的高可信度蛋白873个。其中腺癌与正常黏膜、腺瘤及肝转移灶之间差异显著的蛋白质(定量比值≥2或≤0.5)分别为137、119和77个。结直肠癌与正常黏膜之间的差异蛋白在定位上几乎分布于细胞的各个部位。结论联合应用激光捕获显微切割技术及乙酰化稳定同位素标记定量技术进行结直肠癌蛋白质组学研究,发现的差异蛋白功能上参与多种生物学过程,表明本研究所采取的技术策略有很好的蛋白鉴定覆盖率,可成为研究肿瘤蛋白质组学的有效策略。Abstract: Objective To establish expression profile of differential proteins in colorectal cancers by proteomics technology. Methods Twenty cases of purified cell of colorectal adenocarcinoma,the corresponding normal mucosa,20 cases of villous adenoma and 8 cases of hepatic metastasis were obtained using laser capture microdissection. The differential protein expressions were identified using quantitative proteomic strategy based on acetylation stable isotope labeling. Results Eight hundred and seventy-three kinds of differentially expressed proteins were screened.There were 137,119 and 77 significantly differential proteins in colorectal cancer and normal mucosa or villous adenoma or hepatic metastasis. Conclusion Colorectal cancer related differential proteins covered a wide range of subcellular localizations and involved in many biological processes.It means that the methods have a wide and effective protein coverage and may become an effective strategy for oncoproteomics study.
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