利用蛋白质组学技术研究三氯乙烯刺激肝细胞后蛋白质差异表达  

Study on differential proteomic expression in human liver cells stimulated by trichloroethylene with proteomics

作　　者：刘建军[1] 黄海燕[2] 庄志雄[2] 李习艺[2] 袁建辉[2] 阳帆[1] 卫秦芝[2] 

机构地区：[1]深圳市疾病预防控制中心微生物检验科,518020 [2]深圳市疾病预防控制中心食品毒理科,518020

出　　处：《中华劳动卫生职业病杂志》2005年第6期431-434,共4页Chinese Journal of Industrial Hygiene and Occupational Diseases

基　　金：国家重点基础研究发展规划项目(973)(2002CB512904);国家自然科学基金资助项目(30571557);广东省自然科学基金资助项目(34970;5009153);深圳市科技计划项目(JH200505300493A;200204118)

摘　　要：目的应用双向电泳及串联质谱技术,研究不同剂量三氯乙烯(TCE)诱导L-02肝细胞蛋白质表达谱的差异。方法L-02肝细胞分别用不同剂量TCE和溶剂对照(二甲亚砜)处理后,提取细胞总蛋白,双向凝胶电泳分离蛋白质组成分,银染显色,经ImageMaster2D Platinum5.0软件分组分析比较四组图谱,并对差异蛋白斑点进行基质辅助激光解吸/电离飞行时间串联质谱(MALDI-TOF-TOF-MS)分析鉴定。结果得出差异较明显的蛋白质斑点15个,不同剂量TCE刺激后表达分别上调、下调或消失,提示TCE可引起肝细胞蛋白表达发生改变。通过质谱鉴定和IPI human数据库检索,鉴定了其中9个TCE诱导L-02肝细胞相关的蛋白质。结论这些结果为深入研究TCE毒作用相关的蛋白质及TCE毒作用的分子标志物提供依据,并为阐明TCE毒作用的分子机制提供线索。Objective To explore the differential proteomic expression in human liver cells L-02 induced by different dosages of trichloroethylene（TCE）. Methods Human liver cells L-02 were treated with different concentrations of TCE and the solvent control（dimethylsulfoxide）. The total cellular proteins were separated using 2DE and visualized with silver staining after TCE treatment. The images were analyzed with ImageMaster 2D Platinum 5.0 analysis software. The differentially expressed protein spots were identified by matrix assisted laser desorption ionization-time of flight mass spectrometry（MALDI-TOF-TOF-MS）. Results Fifteen protein spots with significant difference were found, and went upward or downward or disapeared after the stimulation of TCE with different dosages, which indicated that rICE induced the change of the proteomic expression in the liver cells. The mass sprectrmn identification and the IPI human database retrieval were used for identifying 9 proteins related to the L-02 Liver cells induced by TCE. Conclusion The result provides an insight to TCE-related molecular mechanism and which might be useful for further study of the TCE-associated proteins and molecular markers.

关 键 词：三氯乙烯 肝细胞 蛋白质组 电泳 凝胶 电泳 

分 类 号：R131[医药卫生—劳动卫生]