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目的比较永生化食道上皮细胞系和食道鳞状上皮细胞癌(ESCC)细胞系的差异表达细胞骨架相关蛋白图谱,筛选并探讨与ESCC发生发展相关的细胞骨架及相关蛋白。方法采用组分蛋白分离方法提取细胞骨架相关蛋白,利用双向凝胶电泳(2DE)联合MALDI-TOF/TOF串联质谱技术,分析并鉴定永生化食道上皮细胞系NE1及ESCC细胞系HKESC-2中差异表达的细胞骨架及相关蛋白,并用Western blot验证其中感兴趣的差异蛋白在一组食道上皮来源细胞系和ESCC患者肿瘤组织中的表达情况。结果三次独立实验结果表明永生化食道上皮细胞系NE1和ESCC细胞系HKESC-2有52个蛋白点表达差异超过2倍,31个蛋白点经质谱分析鉴定为28种蛋白质,其中16种蛋白质在HKESC-2细胞中表达上调,12种表达下调。Vimentin在所有ESCC细胞系和大部分ESCC肿瘤组织中均呈现高表达,而Annexin II则呈现低表达甚至缺失。结论多种细胞骨架及相关蛋白在ESCC发生发展过程中表达发生变化,某些细胞骨架及相关蛋白的特异性表达模式可能可以作为ESCC诊断的分子标志。
Objective To compare differentially expressed cytoskeleton related proteins between immortalized esophageal epithelial cell lines and esophageal squamous cell carcinoma (ESCC) cell lines and to screen and explore cytoskeleton and related proteins related to the development of ESCC. Methods The cytoskeletal proteins were extracted by protein separation. The differences between immortalized esophageal epithelial cell line NE1 and ESCC cell line HKESC-2 were analyzed and identified by two-dimensional gel electrophoresis (2DE) combined with MALDI-TOF / TOF tandem mass spectrometry The expression of cytoskeleton and related proteins, and Western blot was used to verify the differential proteins of interest in a group of esophageal epithelial cell lines and ESCC patients with tumor tissue. Results The results of three independent experiments showed that the expression of 52 protein spots in NE1 and ESCC cell lines HKESC-2 was more than 2 times higher than that in immortalized esophageal epithelial cell line ESH-ESCC, and 31 protein spots were identified as 28 proteins by mass spectrometry. 16 of them were expressed in HKESC -2 cells were up-regulated, 12 down-regulated. Vimentin is highly expressed in all ESCC cell lines and most of ESCC tumor tissues, while Annexin II shows low or even no expression. Conclusion The expression of many cytoskeletons and related proteins changes during the development of ESCC. The specific expression patterns of some cytoskeletons and related proteins may serve as molecular markers for the diagnosis of ESCC.