SELDI技术分析体外培养不同肝细胞株差异蛋白的表达论文.docVIP

　　SELDI技术分析体外培养不同肝细胞株差异蛋白的表达论文 丁守怡，钱冬萌，牟文凤，闫志勇，宋旭霞，王斌 【关键词】 表面增强激光解吸/离子化/飞行时间质谱；蛋白质陈列分析；肝肿瘤；肿瘤细胞,培养的；差异蛋白 Analysis of different protein expressions in different liver cell strains in vitro using SELDI 【Abstract】 AIM: To examine the differential expressions of proteins in hepatoma cell line (HepG2),hepatoma cell line transfected by HBV （HepG2.2.15）pared al liver cell line(LO2) by using surfaceenhanced laser desorption/ionization timeofflight mass spectrometry (SELDITOFMS), so as to establish a foundation for further studying on the mechanism of hepatoma at the protein level. METHODS: The 3 types of cells above mentioned ployed to detect the differential expressions of proteomes of HepG2, HepG 2.2.15 and LO2. RESULTS: Niyone proteins al liver cell lines, in the segments from Mr 5000 to 15 000, proteins in the HepG2 and HepG2.2.15 shoa cells, the other 5 decreased. Nine proteins in HepG2 ethod in detecting the biological markers in the earlier period of hepatoma. This method is of convenience, high sensitivity, and good reproducibility. The biological tags of tissuespecific proteins a. These tags are also meaningful in screening and identifying signal proteins from serum and tissue specimen in different types of hepatoma. Certain foundation has been established in studying the etiopathogenesis of hepatoma at the level of proteins, as s; tumor cells,.freelRNA的水平并不能真正代表所表达的蛋白质水平，因此，要求对生物功能的执行者蛋白质进行研究. 蛋白质组是指一个基因组、一个细胞或组织或一种生物体所表达的全部蛋白质［1］. 蛋白质组学（Proteomics）是蛋白质组概念的延伸，是在整体水平上研究细胞内蛋白质组成及其活动规律的学科. 蛋白质组学技术为研究肿瘤标志物与肿瘤进展转移研究提供良好的技术平台, 为研究开辟了新的手段和视角. SELDI蛋白质芯片技术是近年来新兴的一种蛋白质组学技术，它具有简单、快捷、灵敏等特点，可以检测分子量在Mr 500~500 000之间的蛋白或多肽，而且所需样本体积小（0.5~400 μL）［2-3］. 我们应用细胞裂解液裂解体外培养的3种细胞（LO2, HepG2, HepG2.2.15），进行了表面增强激光解吸/离子化/飞行时间质谱技术（SELDITOFMS）分析，初步建立了正常肝细胞、肝癌细胞与转染乙肝病毒的肝癌细胞的蛋白表达图谱［4］，发现了一系列（信噪比）差异表达的蛋白，为今后从血清或肝癌组织中筛选标志蛋白提供科学依据. 1材料与方法 1.1材料正常肝细胞株（LO2）购自中科院上海细胞库；肝癌细胞株（HepG2）及携带乙肝病毒的肝癌细胞株（HepG2.2.15）由第四军医大学吴力克主任医师赠送. HPLC水，乙晴，三氟乙酸，白芥子酸（SPA）,TritonX100，尿素，4羟乙基哌嗪乙磺酸（HEPES），表面活性剂（CHAPS）均购自