Knox Bridgett, Wang Yong, Rogers Lora J, Xuan Jiekun, Yu Dianke, Guan Huaijin, Chen Jiwei, Shi Tieliu, Ning Baitang, Kadlubar Susan A
US Food and Drug Administration, National Center for Toxicological Research, Jefferson, Arkansas.
Eye Institute, Affiliated Hospital of Nantong University, Nantong, Jiangsu, China.
Environ Mol Mutagen. 2018 Mar;59(2):134-143. doi: 10.1002/em.22159. Epub 2017 Dec 5.
The transporter associated with antigen processing 2 (TAP2) is involved in the development of multidrug resistance and the etiology of immunological diseases. In this study, we investigated whether the expression of TAP2 can be perturbed by single nucleotide polymorphisms (SNPs) located in 3'-untranslated region (3'-UTR) of the gene via interactions with microRNAs. Using a series of in silico assays, we selected the candidate microRNAs (miRNAs) with the potential to interact with functional SNPs of TAP2. The SNP rs241456-located in the 3'-UTR of TAP2-resides in a potential binding site for hsa-miR-1270 and hsa-miR-620. HEK 293 cells, from a human kidney cell line, were used to characterize the extent of binding of miRNAs to each polymorphic allele of the SNP by a luciferase reporter gene assay. RNA electrophoretic mobility shift assays were used to evaluate the interaction between the miRNAs and each allele sequence of the SNP. We found that hsa-miR-1270 inhibited luciferase activity by binding to the T allele of the SNP in an allele-specific manner. A negative correlation was also found between the expression of hsa-miR-1270 and the T allele of the SNP in kidney tissues. Our findings support the hypothesis that hsa-miR-1270 suppresses the production of TAP2 by binding to this SNP in the 3'-UTR of this gene. Environ. Mol. Mutagen. 59:134-143, 2018. © 2017 Wiley Periodicals, Inc.
抗原加工相关转运体2(TAP2)与多药耐药性的发展及免疫性疾病的病因有关。在本研究中,我们调查了位于该基因3'非翻译区(3'-UTR)的单核苷酸多态性(SNP)是否通过与微小RNA相互作用而干扰TAP2的表达。通过一系列计算机分析,我们筛选出了有可能与TAP2功能性SNP相互作用的候选微小RNA(miRNA)。位于TAP2基因3'-UTR的SNP rs241456存在于hsa-miR-1270和hsa-miR-620的潜在结合位点中。来自人肾细胞系的HEK 293细胞被用于通过荧光素酶报告基因检测来表征miRNA与该SNP各多态性等位基因的结合程度。RNA电泳迁移率变动分析被用于评估miRNA与该SNP各等位基因序列之间的相互作用。我们发现hsa-miR-1270以等位基因特异性方式与该SNP的T等位基因结合,从而抑制荧光素酶活性。在肾组织中还发现hsa-miR-1270的表达与该SNP的T等位基因之间呈负相关。我们的研究结果支持以下假设:hsa-miR-1270通过与该基因3'-UTR中的此SNP结合来抑制TAP2的产生。《环境与分子突变》59:134 - 143,2018年。© 2017威利期刊公司