人鼻咽癌细胞系HONE1的蛋白质双向凝胶电泳分析

目的：研究人鼻咽癌细胞系HONE1的蛋白质表达特征。方法：抽提鼻咽癌细胞系HONE1总蛋白，双向凝胶电泳（2－DE）分离、银杂显色，用凝胶成像仪和PDQUEST软件获取HONE1总蛋白的2－DE图像；根据获得的2－DE凝胶图像，从胶上切取分辨良好、染色较浓的蛋白质点，用胰蛋白酶原位水解，所获酶解肽段经基质辅助激光解吸飞行的时间质谱（MALDI－TOF－MS）测定Mr；以测得肽段Mr为肽质量指纹（PMF），通过Pepldent软件搜索SWISS－PROT和TrEMBL数据库而识别该点对应的蛋白质。结果：建立了双向凝胶电泳分离总蛋白、质谱鉴定银染蛋白质点的方法，并鉴定出人鼻咽癌细胞系HONE1中表达较强的10个点相应的蛋白质。 结论：本研究为建立人鼻咽癌细胞HONE1的2－DE参考图和蛋白质数据库提供了有用的基础性研究资料，为进一步识别鼻咽癌特异性的蛋白质奠定了实验基础。

Two-dimensional gel electrophoresis analysis of proteins from human nasopharyngeal carcinoma cell line HONE1

Purpose:To study the expression characteristics of nasopharyngeal carcinoma cell line at the protein level. Methods:The total proteins of nasopharyngeal carcinoma cell line HONE1 were extracted and separated with two-dimensional gel electrophoresis (2-DE). After gels silver staining, the gel images were acquired by camera and 2-DE analysis software PDQUEST. The distinct and deep-color protein spots were incised from gels and digested by trypsin in-gel, then the matrix-assisted laser desorption/ionization time of flying mass spectrometry (MALDI-TOF-MS) were used for proteins identification. Tryptic peptide mass fingerprints (PMFs) were ultilited to search the SWISS-PROT and TrEMBL database with PepIdent software. Results:The combination of 2D-E and MS method have been established in our lab, 10 proteins in HONE1 cell line were identified. Conclusions:Our results provide fundamental information for constructing a reference map and protein database of human nasopharyngeal carcinoma cell line HONE1 or recognizing the special proteins of nasopharyngeal carcinoma.