转录因子Sp1对胆酸转运蛋白MRP3、MRP4表达的影响  

Influence of transcriptional factor Sp1 on expression of bile acids transporters MRP3 and MRP4

作　　者：陈琨[1] 柴进[1] 封欣婵 程英[1] 张樑君 胡珍[1] 陈兰芳[1] 陈文生[1] 何勇虹[1] 

机构地区：[1]第三军医大学西南医院全军消化病研究所,重庆400038

出　　处：《局解手术学杂志》2016年第7期469-473,共5页Journal of Regional Anatomy and Operative Surgery

基　　金：国家自然基金面上项目(81370560;81170430;81470880)

摘　　要：目的研究胆汁淤积时Sp1对MRP3、MRP4表达的影响,以完善MRP3、MRP4表达调控机制。方法构建Sp1过表达质粒及Sp1 siRNA片段,分别转染HepG2细胞,筛选稳转细胞株。提取细胞总RNA和总蛋白,RT-qPCR检测MRP3、MRP4 mRNA水平的变化,Western blot检测MRP3、MRP4蛋白水平的变化。结果成功构建了Sp1-OE-HepG2和Sp1siRNA-HepG2稳转细胞株。在Sp1-OE-HepG2细胞中,MRP3、MRP4 mRNA和蛋白水平表达均增高,其中MRP3 mRNA水平表达增高2.8倍,MRP3蛋白水平表达增高3.0倍;MRP4 mRNA和蛋白水平分别增高3.2倍和2.5倍;而在Sp1 siRNA-HepG2细胞中,MRP3、MRP4则表达下降,其中MRP3 mRNA表达降低52%,MRP4 mRNA表达降低58%,MRP3蛋白表达降低57%,MRP4蛋白表达降低60%。结论转录因子Sp1能够调控胆酸转运蛋白MRP3、MRP4的表达。Objective To investigate the influence of transcriptional factor Sp1 on expression of bile acids transporters MRP3 and MRP4 so as to perfect the regulatory mechanism of MRP3 and MRP4 expression. Methods Transformed Sp1-overexpression and Sp1 siRNA plasmids to HepG2 cell and obtained the stably cells line. Then the expression levels of bile acids transporters MRP3 and MRP4 were measured by RT-qPCR,and the change of protein levels were detected by Western blot. Results The stably cells line Sp1-OE-HepG2 and Sp1 siRNAHepG2 were successfully transformed. The mRNA expression and protein levels of MRP3 and MRP4 were significantly increased in Sp1-OEHepG2 cells,among which the mRNA expression of MRP3 mRNA increased 2. 8 times,the protein levels of MRP3 increased 3. 0 times,and the mRNA expression and protein levels of MRP4 increased 3. 2 times and 2. 5 times respectively. Conversely,the mRNA expression and protein levels of MRP3 and MRP4 were decreased in Sp1 siRNA-HepG2 cells,among which the mRNA expression of MRP3 mRNA decreased 52%,the mRNA expression of MRP4 mRNA decreased 58%,the protein levels of MRP3 decreased 57%,and the protein levels of MRP4 decreased 60%. Conclusion Transcriptional factor Sp1 could regulate the expression of bile acids transporters MRP3 and MRP4 in HepG2 cells.

关 键 词：胆汁淤积 转录因子 SP1 MRP3 MRP4 

分 类 号：R34[医药卫生—基础医学] R575.5