导向激光捕获显微切割纯化的贲门腺癌细胞蛋白质表达图谱的建立  

Establishment of protein expression profile of navigated laser capture microdissected human gastric cardia adenocarcinoma

作　　者：程妍[1] 张军[1] 刘冬[1] 伍洁[2] 宋亚华[1] 龚均[1] 

机构地区：[1]西安交通大学第二附属医院消化内科,西安710004 [2]西安交通大学第二附属医院病理科

出　　处：《山西医科大学学报》2015年第8期740-746,共7页Journal of Shanxi Medical University

基　　金：国家自然科学基金资助项目(30470785)

摘　　要：目的建立高分辨率的贲门腺癌细胞双向凝胶电泳图谱,为构建贲门腺癌的蛋白质表达数据库奠定基础。方法应用导向激光捕获显微切割技术(NLCM)获取贲门腺癌细胞,应用双向凝胶电泳(2-DE)技术对贲门腺癌细胞蛋白进行分析,建立贲门腺癌细胞的蛋白质表达图谱,并通过基质辅助激光解析电离飞行时间质谱肽质量指纹技术(MALDI-TOF MS)和数据库对部分蛋白质点进行鉴定。结果获得了背景清晰、分辨率高、重复性好的NLCM分离的贲门腺癌细胞双向凝胶电泳图谱;鉴定出了72个贲门腺癌相关蛋白质;蛋白质大部分位于细胞质和线粒体,其次位于细胞膜、细胞核、细胞骨架、内质网、高尔基体、核糖体等。蛋白质涉及细胞代谢、增殖和分化、RNA转录和翻译、信号转导、氧化还原、蛋白质分子伴侣等。结论首次建立了分辨率高、重复性好的导向激光捕获显微切割分离的贲门腺癌细胞的双向凝胶电泳图谱,并鉴定出了72个贲门腺癌相关蛋白,为进一步构建贲门腺癌的蛋白质组学数据库奠定了基础。Objective To obtain the two-dimensional gel electrophoresis （2-DE） patterns of human gastric cardiac adenocarcinoma （GCA） cells captured by navigated laser capture microdissection（NLCM） with high resolution and reproducibility.Methods Navigated LCM was used to obtain the malignant gastric cardiac cells from surgical specimens of human GCA.The proteins extracted from these cells were then separated by 2-DE.Differential protein spots were identified by peptide mass fingerprint（PMF） based on matrixassisted laser desorption/ionization time-of-flight mass spectrometry （MALDI-TOF MS） and database searching.Results ①The 2-DE patterns of human GCA cells captured by NLCM were successfully obtained with high resolution and reproducibility.②With mass spectrometry technology（peptide mass fingerprint）,72 protein spots were successfully identified.The identified proteins of GCA have been classified in terms of their cellular localization and physiological function using information from SWISS-PROT and NCBI websites.③The protein was mostly located in the cytoplasm and mitochondrion,followed by membrane,nucleus,reticulum,cytoskeleton,ribosomal subunit and Golgi apparatus.The proteins were involved in the metabolic progress,cell proliferation and differentiation,transcription and translation,signal transduction,chaperones and oxidation-reduction.Conclusion The 2-DE pattern of human GCA is obtained with high resolution and reproducibility and 72 proteins of GCA are identified,which may be helpful to construct the database.

关 键 词：贲门腺癌 激光捕获显微切割 蛋白质组 分子标志物 

分 类 号：R735.2[医药卫生—肿瘤]