C/EBP β对APP基因的表达调控作用的研究  

Effects of C/EBP β on APP Gene Expression

作　　者：柴娟 李永玲[2] 芦晓红[2] 张茜[2] 马琳 Huichen Wang 孙涛 崔建奇[1,2] 

机构地区：[1]宁夏颅脑疾病重点实验室,省部共建国家重点实验室培育基地,银川750004 [2]宁夏医科大学基础医学院,银川750004 [3]美国艾莫里大学Winship肿瘤研究所,放射肿瘤学研究室

出　　处：《中国细胞生物学学报》2015年第4期500-507,共8页Chinese Journal of Cell Biology

基　　金：国家重点基础研究发展计划(973计划)(批准号:2012CB722408);国家自然科学基金(批准号:81260197)资助的课题~~

摘　　要：该研究成功地构建了C/EBP β过表达慢病毒载体,在体外人胚肾细胞(HEK293FT)进行病毒包装并感染小鼠海马神经元细胞(HT22)。通过荧光素酶报告基因实验(luciferase assay)检测C/EBPβ对淀粉样前体蛋白(amyloid precursor protein,APP)启动子活性的影响;通过实时荧光定量PCR(Q-PCR)来检测C/EBPβ对APP和Sp1在转录水平上的表达;通过蛋白免疫印迹分析(Western blot assay)检测C/EBPβ对APP和Sp1蛋白表达的作用。萤火虫荧光素酶分析结果显示,C/EBPβ对APP启动子的表达有正调控作用;Western blot和Q-PCR分析的结果表明,C/EBPβ对APP和Sp1基因的表达有正调控作用。C/EBPβ对APP基因表达的调控作用的机制可能在于C/EBPβ上调了内源性转录因子Sp1的基因表达,而Sp1基因表达的增强直接导致了APP基因表达的上调。The C/EBP β lentivirus expression vector was successfully constructed and the viron was packaged by virus packaging kit in HEK293 FT cells. Rat hippocampus neuron（HT22） was infected with the packaged viron for the study. The luciferase assay was performed to evaluate the effects of C/EBP β on APP promoter activities; the effects of C/EBP β on amyloid precursor protein（APP） and Sp1 gene expression were estimated with Q-PCR and Western blot. The results of luciferase assay demonstrated that C/EBP β could up-regulate APP promoter activities, and the results of Q-PCR and Western blot confirmed that C/EBP β could increase APP and Sp1 gene expressions both in transcriptional and translational levels. The possible mechanism for C/EBP β to upregulate APP gene expression may lie in C/EBP β facilitating the endogenous Sp1 gene expression, and the increasing expression of Sp1 directly promotes the APP gene expression.

关 键 词：C/EBP β 淀粉样前体蛋白 阿尔茨海默病 SP1 基因表达调控 

分 类 号：R373[医药卫生—病原生物学]