小鼠肝细胞胰岛素与表皮生长因子信号转导磷蛋白质组的对比分析(英文)  

Dynamic phosphoproteomics of insulin and epidermal growth factor signaling in mouse hepatocytes

作　　者：刘友平[1] 姚富丽[1] 丁慧荣[1] 李洪[1] 

机构地区：[1]泸州医学院生物化学教研室,四川省泸州市646000

出　　处：《中国组织工程研究与临床康复》2008年第42期8380-8383,共4页Journal of Clinical Rehabilitative Tissue Engineering Research

基　　金："细胞信号转导的复杂性与计算机模拟":国家自然科学基金资助(70071040)~~

摘　　要：背景:表皮生长因子和胰岛素都主要通过PI3K通路和MAPK通路进行信号转导,但各自的生理功能不同。目的:从细胞整体水平对比分析小鼠肝细胞内胰岛素与表皮生长因子信号转导磷蛋白质组的动力学行为差别,以此找出二者关键性的信号蛋白。设计、时间及地点:随机分组设计的对比观察实验,于2005-07/2006-04在泸州医学院分子生物学实验室完成。材料:实验用肝细胞来源于昆明种封闭群乳小鼠。方法:选取细胞长势相当的肝细胞,采用放射性同位素(32P)对其进行标记,标记后按随机数字表法分为3组,空白对照组、表皮生长因子刺激组(给予10μg/L表皮生长因子)及胰岛素刺激组(给予100nmol/L胰岛素)。主要观察指标:表皮生长因子、胰岛素分别作用0,5,20,60,120min,采用双向电泳技术分析比较表皮生长因子与胰岛素分别介导的磷酸化蛋白质组的动力学行为,即磷酸化水平。结果:参与两者信号转导的磷蛋白质种类没有多大差异,大多数信号磷蛋白磷酸化水平的动力学行为也表现为一致性,但有4个蛋白点其磷酸化水平随时间变化趋势明显不同。结论:胰岛素与表皮生长因子在同一细胞内参与信号转导的个别信号蛋白其磷酸化水平的动力学行为差别较大,估计这些蛋白就是导致表皮生长因子与胰岛素最终生物学活性的关键蛋白。BACKGROUND： Both epidermal growth factor （EGF） and insulin transfer their signals into cells by two primary signal transduction pathways, including phosphatidylinositol 3-kinase （PI3K） pathway and mitogen activated protein kinases （MAPK） pathway. But they have different physiological functions. OBJECTIVE： To comparatively assay the dynamic behaviors of phosphoproteomes between EGF and insulin signal transductions in mouse hepatocytes and find key signal proteins. DESIGN, TIME AND SETTING： Randomized grouping controlled observation experiment was performed in the laboratory of Molecular Biology, Luzhou Medical College between July 2005 and April 2006. MATERIALS： Hepatocytes were from Kunming mice of closed population. METHODS： The primarily cultured mouse hepatocytes were labeled with 32p isotope and then randomly divided into three groups： control, EGF-stimulated （received 10μ g/L EGF）, and insulin-stimulated （received 100 nmol/L insulin） groups. MAIN OUTCOME MEASURES： After mouse hepatocytes were treated with EGF and insulin for 0, 5, 20, 60 and 120 minutes, the dynamic behaviors of phosphoproteomes（i.e, phosphorylated level） between EGF and insulin signal transductions were comparatively analyzed by two-dimensional electrophoresis method. RESULTS： The categories of all phosphorylated proteins between EGF and insulin-stimulated phosphoproteomes had no apparent difference. The dynamic behaviors of phosphoproteomes of most proteins during EGF signal transduction are parallel with thoseduring insulin stimulation, except the dynamic behaviors of 4 proteins are different significantly. CONCLUSION： Aforementioned 4 phosphorylated proteins were most probably the key members that could distinguish between two signal transduction pathways or networks, and determined their major physiological functions respectively.

关 键 词：胰岛素 表皮生长因子 磷酸化蛋白质 信号转导系统 动力学行为 

分 类 号：R735.2[医药卫生—肿瘤] R972[医药卫生—临床医学]