双向凝胶电泳-飞行时间质谱技术鉴定食管上皮细胞癌变时14-3-3 protein ε、S100A9的表达

摘要] 目的：分析食管鳞癌和正常食管上皮细胞蛋白质的表达差异，获取鉴别两者的分子标志物。方法：通过激光捕获显微切割技术分离ESCC肿瘤细胞和癌旁上皮细胞，通过双向电泳和质谱技术鉴定表达异常的蛋白，并通过蛋白免疫印记证实部分差异蛋白的表达。结果：建立了食管癌组织和正常食管上皮蛋白的双向凝胶电泳图谱，通过质谱技术鉴定出14-3-3 protein ε、S100A9等蛋白在食管癌变时差异表达，蛋白印记结果证实14-3-3 protein ε、S100A9的表达量在食管癌变时分别上调和下调。结论：激光捕获显微切割是蛋白质组研究中的一个突破性的技术，可以有效地解决组织异质性的问题；本实验检测到的差异蛋白例如14-3-3 protein ε、S100A9可能成为鉴别食管癌组织和正常食管上皮特异性的分子标记物。

Differential Expression of 14-3-3 protein ε,S100A9,in Cancerization of Human Esophageal Epithelial Cells

ABSTRACT：Objective：To search for the biomarkers for differential diagnosis between human esophageal squamous cell carcinoma cells and normal esophageal epithelial cells. Methods：Laser capture microdissection was used to separate human esophageal squamous cell carcinoma cells and normal esophageal epithelial cells, Proteome alterations were observed using two dimensional polyacrylamide gel electrophoresis(2-DE) technique and electrospray ionisation tandem mass spectrometry．Westernbloting was used to detect the different protein expressions in ESCC and esophageal epithelial cells, Results : The 2-DE patterns of human esophageal squamous cell carcinoma cells and normal esophageal epithelial cells were established. 14-3-3 protein ε、S100A9 were identified differentially expressed between ESCC and esophageal epithelial cells. Conclusion ：Laser capture microdissection is a key technique in proteomic study which can effectively resolve the problem of irrelevant tissues．14-3-3 protein ε、S100A9 may serve as abiomarker for differential diagnosis of human esophageal squamous cell carcinoma cells and normal esophageal epithelial cells.