氢醌刺激后人肝细胞蛋白质谱表达的变化  

Differential proteomic expression in human liver cells stimulated by hydroquinone

作　　者：鞠莉[1] 张淑芝[2] 赵苒[1] 姚耿东[1] 

机构地区：[1]浙江大学环境研究所,杭州310031 [2]杭州市疾病预防控制中心

出　　处：《中华劳动卫生职业病杂志》2006年第11期658-661,共4页Chinese Journal of Industrial Hygiene and Occupational Diseases

摘　　要：目的研究氧醌(HQ)刺激后L-02型人肝细胞蛋白质谱的表达变化。方法传代培养的L-02型肝细胞分为空白对照组和染毒处理组。处理组经HQ(1×10-4mol／L)染毒培养24 h,对照组不做处理。提取蛋白同时进行双向凝胶电泳,图谱经PDQuest软件分析寻找差异点。实验独立进行3次,每次处理组和对照组各设3块胶作为组内重复,切取重复性较好的差异点做MALDI-TOF质谱鉴定,得到PMF通过Mascot在美国国家生物信息检索中心(NCBI)人类蛋白质数据库搜索。结果每块胶检测到约1 000个蛋白点,对照组和处理组凝胶匹配率组内达90％以上,组间80％以上,3次实验各发现17、18、24个蛋白点表达改变,变化倍数≥2。重复性较好的4个蛋白点经MALDI-TOF质谱鉴定为Rho GDP解离抑制蛋白、6-磷酸葡萄糖内酯酶、erbB3结合蛋白EBP1和核纤层蛋白,其功能与信号转导、细胞骨架及能量代谢有关。结论1×10-4mol／L的HQ处理24 h可引起L-02型人肝细胞蛋白质谱表达改变。Objective To explore the differential proleomic expression in human liver cells L-02 after exposure to hydroquinone （HQ）. Methods Subcultured L-02 cells were treated by HQ for 24 h at a 1×10^-4 mol/L concentration and a blank group was set as the control, Immediately after the treatment, total cellular proteins were extracted and separated by 2-DE, and the images were analyzed by PDQuest software. The experiment was totally repeated 3 times with 3 repetitions for each group every, time. The well repeated spots were identified by MALDI-TOF MS and then searched in NCBI human protein database with Mascot. Results About 1 000 spots per gel were found. Compared with the control group, 17,18 and 24 spots were significantly altered in 3 separate experiments. The 4 well repeated spots were identified by MALDI-TOF MS as Rho GDP dissociation inhibitor GDI alpha, 6-phosphogluconolactonase, erbB3 binding protein EBP1 and lamin A/C, isoform 1 precursor. They were involved in cell skeleton, signal transduction and energy metabolization in functional classification. Conclusion HQ can change the protein expression in liver cells, which provides clues for exploring the toxic mechanism.

关 键 词：氢醌 肝细胞 双向凝胶电泳 质谱 蛋白质组 

分 类 号：R114[医药卫生—卫生毒理学]