Functional analysis of the polymorphism -211C>T in the regulatory region of the human ABCC3 gene
Functional analysis of the polymorphism -211C>T in the regulatory region of the human ABCC3 gene
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DOI：
10.1016/j.lfs.2007.01.023
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发表时间：
2007-03-27
期刊：
影响因子：
6.1
通讯作者：
Koening, Joerg
中科院分区：
文献类型：
--
作者：
Gradhand, Ulrike;Tegude, Heike;Koening, Joerg
多药耐药蛋白3 （MRP3/基因符号：ABCC3）是一种atp依赖的外排泵，介导内源性葡萄糖醛酸盐和缀合药物代谢物跨细胞膜的转运。在人类中，ABCC3 mRNA的肝脏表达似乎受到ABCC3基因启动子-211位置C > T多态性的影响。本研究的目的是探讨这种SNP如何影响MRP3表达的可能机制。在翻译起始位点上游0.5、1.1、4.4和8.1 kb的启动子荧光素酶报告基因构建物与-211位置的胞嘧啶或胸腺嘧啶克隆，并转染到HepG2、Caco-2和LS174T细胞中。报告基因的活性取决于启动子序列的长度，但有趣的是，与-211位的核苷酸无关。将FTF cDNA（胎蛋白转录因子）与-211多态性附近的元件共转染，除了独立于-211 SNP的0.5 kb片段外，所有构建体的启动子活性都增加了。综上所述，我们没有发现-211 C > T ABCC3启动子多态性对基础或FTF诱导的报告基因活性有任何影响。其他组织特异性机制是否揭示了该SNP对MRP3体内调控的影响仍有待确定。(c) 2007爱思唯尔公司版权所有。
The multidrug resistance protein 3 (MRP3/gene symbol: ABCC3) is an ATP-dependent efflux pump mediating the transport of endogenous glucuronides and conjugated drug metabolites across cell membranes. In humans the hepatic expression of ABCC3 mRNA seems to be influenced by the polymorphism C > T at the position -211 in the promoter of the ABCC3 gene. The aim of this study was to investigate the possible mechanisms of how this SNP influences the MRP3 expression. Promoter luciferase reporter gene constructs representing 0.5, 1.1, 4.4, and 8.1 kb upstream of the translational start site were cloned with cytosine or thymine at position -211 and transfected into HepG2, Caco-2, and LS174T cells. Reporter gene activity was dependent on the length of the promoter sequence but interestingly not on the nucleotide at position -211. Cotransfection with FTF cDNA (Fetoprotein Transcription Factor) binding to elements near the -211 polymorphism increased promoter activity in all constructs except the 0.5 kb fragment also independently of the -211 SNP. Taken together, we did not find any influence of the -211 C > T ABCC3 promoter polymorphism on either the basal or the FTF induced reporter gene activity. Whether other tissue specific mechanisms reveal an impact of this SNP on the in vivo regulation of MRP3 remains to be determined. (c) 2007 Elsevier Inc. All rights reserved.