2D differential in-gel electrophoresis for the identification of esophageal scans cell cancer-specific protein markers
2D differential in-gel electrophoresis for the identification of esophageal scans cell cancer-specific protein markers
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DOI：
10.1074/mcp.m100015-mcp200
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发表时间：
2002-02-01
影响因子：
7
通讯作者：
Zhao, YM
中科院分区：
文献类型：
--
作者：
Zhou, G;Li, HM;Zhao, YM
差分凝胶电泳（DIGE）是一种新兴的蛋白质组学分析技术，可以提高传统二维凝胶电泳的可重复性。在DIGE中，两个蛋白池分别用1-（5-羧基戊基）-1′-丙基碳氰卤化物（Cy3） n -羟基琥珀酰亚酰基酯和1-（5-羧基戊基）-1′-甲基蓝二碳氰卤化物（Cy5） n -羟基琥珀酰亚酰基酯荧光染料标记。标记的蛋白质在相同的二维凝胶中混合和分离。2D DIGE用于量化激光捕获微解剖获得的食管癌细胞与正常上皮细胞之间蛋白质表达的差异，并定义癌症特异性和正常特异性蛋白质标记物。对25万个癌细胞和正常细胞的蛋白质裂解物的二维图像进行分析，发现癌细胞裂解物中有1038个蛋白质斑点，正常细胞裂解物中有1088个蛋白质斑点。在检测到的蛋白中，癌细胞中有58个位点被>上调3倍，107个位点被>下调3倍。除了先前发现的膜联蛋白I下调外，肿瘤排斥抗原（gp96）在食管鳞状细胞癌中也被发现上调。使用2D DIGE结合质谱技术对激光捕获显微解剖的患者匹配的癌细胞和正常细胞之间的蛋白质表达进行全局定量，是癌症进展分子表征和癌症特异性蛋白质标记物鉴定的有力工具。中国生物医学工程学报（英文版）；2006；
The reproducibility of conventional two-dimensional (2D) gel electrophoresis can be improved using differential in-gel electrophoresis (DIGE), a new emerging technology for proteomic analysis. In DIGE, two pools of proteins are labeled with 1-(5-carboxypentyl)-1'-propylindocarbocyanine halide (Cy3) N-hydroxy-succinimidyl ester and 1-(5carboxypentyl)-1'-methylindodi-carbocyanine halide (Cy5) N-hydroxysuccinimidyl ester fluorescent dyes, respectively. The labeled proteins are mixed and separated in the same 2D gel. 2D DIGE was applied to quantify the differences in protein expression between laser capture microdissection-procured esophageal carcinoma cells and normal epithelial cells and to define cancer-specific and normal-specific protein markers. Analysis of the 2D images from protein lysates of similar to 250,000 cancer cells and normal cells identified 1038 protein spots in cancer cell lysates and 1088 protein spots in normal cell lysates. Of the detected proteins, 58 spots were up-regulated by >3-fold and 107 were down-regulated by >3-fold in cancer cells. In addition to previously identified down-regulated protein annexin I, tumor rejection antigen (gp96) was found up-regulated in esophageal squamous cell cancer. Global quantification of protein expression between laser capture-microdissected patient-matched cancer cells and normal cells using 2D DIGE in combination with mass spectrometry is a powerful tool for the molecular characterization of cancer progression and identification of cancer-specific protein markers. Molecular & Cellular Proteomics 1:117-124, 2002.