The effects of ABCB1 3′-untranslated region variants on mRNA stability
The effects of ABCB1 3′-untranslated region variants on mRNA stability
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DOI：
10.1124/dmd.107.017087
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发表时间：
2008-01-01
影响因子：
3.9
通讯作者：
Kroetz, Deanna L.
中科院分区：
文献类型：
--
作者：
Gow, Jason M.;Chinn, Leslie W.;Kroetz, Deanna L.
编码P-糖蛋白（P-gp）的ABCB 1的遗传变异是药物应答个体间变异的潜在原因。大量的研究集中在编码区变异对P-gp表达和功能的影响，而非编码区变异的研究很少。3 '-非翻译区（UTR）通过与mRNA降解机制的RNA-蛋白质相互作用来调节mRNA水平或稳定性。mRNA稳定性是控制ABCB 1 mRNA表达的关键调控步骤，最终影响P-gp水平和功能。我们假设ABCB 1 3 '-UTR多态性通过破坏RNA-蛋白质相互作用来改变mRNA的稳定性。对一组不同种族的DNA样品进行测序以鉴定3 '-UTR多态性并确定等位基因频率。三种最常见的变体，沿着参考ABCB 1，在细胞中稳定表达，以测量mRNA半衰期。ABCB 1参比品在HEK 293细胞中的计算半衰期为9.4 +/- 1.3 h，与3 '-UTR变体的估计半衰期相似。在携带3 '-UTR变体和参考等位基因的淋巴母细胞系中，内源性ABCB 1 mRNA的衰减相似。尽管检查的ABCB 1 3 '-UTR变体对ABCB 1 mRNA稳定性没有影响，但这些数据代表了确定UTR遗传变异对ABCB 1 mRNA水平影响的首次尝试之一。
Genetic variation in ABCB1, encoding P-glycoprotein (P-gp), is a potential cause of interindividual variation in drug response. Numerous studies have focused on the effects of coding region variants on P-gp expression and function, whereas few noncoding region variants have been investigated. The 3'-untranslated region (UTR) regulates mRNA levels or stability via RNA-protein interactions with mRNA degradation machinery. mRNA stability is a key regulatory step controlling ABCB1 mRNA expression that ultimately affects P-gp levels and function. We hypothesized that ABCB1 3'-UTR polymorphisms alter mRNA stability by disrupting RNA-protein interactions. An ethnically diverse panel of DNA samples was sequenced to identify 3'-UTR polymorphisms and determine allele frequencies. The three most common variants, along with reference ABCB1, were stably expressed in cells in order to measure mRNA half-life. The calculated half-life for ABCB1 reference in HEK293 cells was 9.4 +/- 1.3 h and was similar to that estimated for the 3'-UTR variants. Endogenous ABCB1 mRNA decay was similar in lymphoblastoid cell lines carrying 3'-UTR variant and reference alleles. Although the examined ABCB1 3'-UTR variants have no effect on ABCB1 mRNA stability, these data represent one of the first attempts to determine the influence of genetic variation in UTRs on ABCB1 mRNA levels.