D-葡萄糖对载脂蛋白A5基因表达的影响及其机制研究  

D-glucose affects expression of apolipoprotein A5 and relevant mechanism

作　　者：张葵[1] 陈娟[1] 魏红霞[1] 李雷[1] 宋宏岩[1] 顾光煜[1] 丁福霖[1] 钱世宁[1] 

机构地区：[1]南京大学医学院附属鼓楼医院检验科,南京210008

出　　处：《临床检验杂志》2013年第8期592-595,共4页Chinese Journal of Clinical Laboratory Science

基　　金：南京市科技发展重点项目(ZKX09030)

摘　　要：目的探讨D-葡萄糖及转录因子USF1、USF2对载脂蛋白A5(apo A5)基因表达的影响及其作用机制。方法体外培养人原代肝癌细胞,分别以不同浓度D-葡萄糖(0、5、25 mmol/L)与人原代肝癌细胞作用48 h,荧光实时定量RT-PCR和western blot分别检测人原代肝癌细胞apo A5、USF1和USF2 mRNA和蛋白质表达水平;染色质免疫沉淀(ChIP)技术分析USF1/USF2与apo A5启动子的结合情况。结果 D-葡萄糖呈浓度依赖性增加apo A5基因表达,D-葡萄糖不改变USF1、USF2 mRNA与蛋白质在人原代肝癌细胞中的表达水平(P>0.05);ChIP结果证实,D-葡萄糖可增加转录因子USF1/USF2与apo A5启动子的结合(P<0.05)。结论在D-葡萄糖作用下,转录因子USF1/2可在转录水平调节人原代肝癌细胞中apo A5表达。Objective To investigate the effects of D-glucose,upstream simulatory factor（USF） 1 and 2 on the expression of apolipoprotein A5（apo A5） and the possible mechanism.Methods Primary human hepatoma cells were cultured and incubated with serially diluted D-glucose（0,5,25 mmol / L） for 48 h.The alterations of apo A5,USF1 and USF2 were measured by RT-PCR and western blot analysis at mRNA and protein level respectively.The binding of USF1 /2 to apo A5 promoter was identified by chromatin immunoprecipitation assay.Results D-glucose increased the expression of apo A5,gene in a concentration-dependent manner,while neither the mRNA nor protein levels of both USF1 and USF2 was regulated by incubating primary human hepatoma cells with D-glucose（P 0.05）.Chromatin immunoprecipitation assays showed that D-glucose increased the transcription factor USF1 /2 at the apo A5 gene promoter region（P 0.05）.Conclusion Transcription factor USF1 /2 may upregulate the mRNA and protein levels of apo A5 with the involving effects of D-glucose in primary human hepatoma cells.

关 键 词：载脂蛋白A5 启动子 转录因子 

分 类 号：R735.7[医药卫生—肿瘤]