MicroRNA regulation of expression of the cystic fibrosis transmembrane conductance regulator gene.
MicroRNA regulation of expression of the cystic fibrosis transmembrane conductance regulator gene.
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DOI：
10.1042/bj20110672
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发表时间：
2011-08-15
期刊：
影响因子：
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通讯作者：
Harris A
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文献类型：
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作者：
Gillen AE;Gosalia N;Leir SH;Harris A
囊性纤维化跨膜传导调节因子(CFTR)基因表现出复杂的时空表达模式，受与基本启动子相互作用的多个顺式作用元件控制。尽管在理解这些基因组元件方面已经取得了重大进展，但还没有关于miRNAs(MicroRNAs)对CFTRs转录后调控的报道。在本研究中，我们发现了两个miRNAs，hsa-miR-145和hsa-miR-494，它们通过直接靶向CFTR3‘UTR(非翻译区)上的离散位点来调节CFTR的表达。我们发现至少有12个miRNAs能够抑制Caco-2细胞内源性CFTRmRNA的表达。其中10个还抑制了一个或多个细胞系中含有CFTR3‘UTR的报告结构的表达，5个抑制了Caco-2细胞中内源性CFTR蛋白的表达。此外，至少有三种基因在原代人呼吸道上皮细胞中表达，与肠道细胞系相比，CFTR的表达保持在较低的水平。其中三个针对cftr的miRNAs，hsa-miR-384，hsa-miR-494和hsamiR-1246，也抑制了携带Na+-K+-Cl−共转运体SLc12a2[溶质载体家族12(Na+-K+-ClmiRNAs)，成员2]3‘非编码区的报告基因的表达，这表明这些miRNAs可能在调节上皮细胞中氯的运输中发挥更普遍的作用。
The CFTR (cystic fibrosis transmembrane conductance regulator) gene shows a complex temporal and spatial pattern of expression that is controlled by multiple cis-acting elements interacting with the basal promoter. Although significant progress has been made towards understanding these genomic elements, there have been no reports of post-transcriptional regulation of CFTR by miRNAs (microRNAs). In the present study, we identify two miRNAs, hsa-miR-145 and hsa-miR-494, which regulate CFTR expression by directly targeting discrete sites in the CFTR 3′ UTR (untranslated region). We show that at least 12 miRNAs are capable of repressing endogenous CFTR mRNA expression in the Caco-2 cell line. Ten of these also inhibit expression of a reporter construct containing the CFTR 3′ UTR in one or more cell lines, and five repress endogenous CFTR protein expression in Caco-2 cells. Moreover, at least three are expressed in primary human airway epithelial cells, where CFTR expression is maintained at low levels in comparison with intestinal cell lines. Three of the miRNAs that target CFTR, hsa-miR-384, hsa-miR-494 and hsamiR-1246, also inhibit expression of a reporter carrying the Na+ – K+ –Cl− co-transporter SLC12A2 [solute carrier family 12 (Na+ – K+ –Cl− transporters), member 2] 3′ UTR, suggesting that these miRNAs may play a more general role in regulating chloride transport in epithelial cells.