The Center of Gerontology and Geriatrics, West China Hospital, Sichuan University, Chengdu, China.
National Clinical Research Center of Geriatrics, West China Hospital, Sichuan University, Chengdu, China.
J Cell Mol Med. 2021 Apr;25(8):3714-3723. doi: 10.1111/jcmm.15976. Epub 2021 Feb 26.
Genetic variations in the 3'UTR of mRNAs as well as sequences of microRNAs (miRNAs) and long non-coding RNAs (lncRNAs) can affect gene expression by interfering with the binding between them. In this study, we investigated the role of the following polymorphisms in the risk of hypertension: the 774T > C (rs17337023) polymorphism located in the EGFR 3' untranslated region (3'UTR), the rs884225 polymorphism located in the sequence of miR-214, and the single nucleotide polymorphisms (SNPs) rs325797437, rs344501106, rs81286029 and rs318656749 located in the promoter of lncRNA MEG3. Taqman genotyping assays and haplotype analysis tools were used to measure the MEG3 haplotypes and the rs17337023 and rs884225 polymorphisms genotypes. The relationship between MEG3, miR-214 and EGFR was validated using computational analysis and luciferase assays. Unlike other polymorphisms, only patients grouped according to their rs884225 genotypes exhibited varied EGFR mRNA and protein levels, which indicated that the rs884225 genotype is associated with the expression of EGFR mRNA and protein levels. MiR-214 was confirmed to bind to MEG3 and 3'UTR of EGFR by showing that the transfection of exogenous miR-214 significantly down-regulated the luciferase activity of A549 and H460 cells transfected with wild-type MEG3 or wild-type EGFR 3' UTR. Additionally, MEG3 overexpression inhibited miR-214 expression while elevating the EGFR mRNA and protein expressions. Meanwhile, MEG3 down-regulation demonstrated an opposite result, thus establishing the MEG3/miR-214/EGRF signalling pathway. Our study confirmed that the T > C substitution of rs884225 polymorphism located in miR-214 binding site in the 3'UTR of EGFR is associated with increased risk of primary hypertension.
mRNA 的 3'UTR 中的遗传变异以及 microRNAs (miRNAs) 和长非编码 RNA (lncRNAs) 的序列可以通过干扰它们之间的结合来影响基因表达。在这项研究中，我们研究了以下多态性在高血压风险中的作用：位于 EGFR 3'非翻译区 (3'UTR) 的 774T>C(rs17337023) 多态性、位于 miR-214 序列中的 rs884225 多态性，以及位于 lncRNA MEG3 启动子中的单核苷酸多态性 (SNP) rs325797437、rs344501106、rs81286029 和 rs318656749。Taqman 基因分型检测和单倍型分析工具用于测量 MEG3 单倍型和 rs17337023 和 rs884225 多态性基因型。使用计算分析和荧光素酶检测验证了 MEG3、miR-214 和 EGFR 之间的关系。与其他多态性不同，只有根据 rs884225 基因型分组的患者表现出不同的 EGFR mRNA 和蛋白水平，这表明 rs884225 基因型与 EGFR mRNA 和蛋白水平的表达有关。通过显示转染外源性 miR-214 可显著下调转染野生型 MEG3 或野生型 EGFR 3'UTR 的 A549 和 H460 细胞的荧光素酶活性，证实了 miR-214 与 MEG3 和 EGFR 3'UTR 的结合。此外，MEG3 过表达抑制 miR-214 表达，同时升高 EGFR mRNA 和蛋白表达。同时，MEG3 下调显示出相反的结果，从而建立了 MEG3/miR-214/EGRF 信号通路。我们的研究证实，位于 EGFR 3'UTR 中 miR-214 结合位点的 rs884225 多态性的 T>C 取代与原发性高血压风险增加有关。
