Department of Neurology, the Affiliated Yueqing Hospital of Wenzhou Medical University, 338 Qingyuan Road, Yueqing, Wenzhou, Zhejiang Province, China.
Department of Encephalopathy, Zhejiang Rehabilitation Medical Center, 2828 Binsheng Road, Binjiang District, Hangzhou, Zhejiang Province, China.
J Mol Neurosci. 2020 Jun;70(6):878-886. doi: 10.1007/s12031-020-01513-y. Epub 2020 Mar 2.
The purpose of this study is to investigate the correlation between single-nucleotide polymorphism (SNP) at the 3' end of the untranslated region (UTR) of Sirtuin 2 (SIRT2) gene and the risk of developing Alzheimer's disease (AD), and to explore its underlying mechanisms. In total, 260 patients with AD and 260 healthy controls were recruited in this study. The genotype of rs2015 and rs2241703 loci of the SIRT2 gene was analyzed by Sanger sequencing for all participants. Quantitative real-time Polymerase chain reaction (qRT-PCR) was used to analyze microRNAs (miRNAs) and SIRT2 mRNA levels. Western blotting was used to analyze the expression level of SIRT2 protein. The dual luciferase reporter gene assay and cell transfection were performed to examine the role of miRNAs in regulating SIRT2 expression. Carriers of the SIRT2 gene rs2015 locus A allele were 0.69 times less likely to develop AD than the carriers of the C allele (95% confidence interval (CI): 0.59-0.80, p < 0.01). The carriers of the SIRT2 gene rs2241703 locus A allele were 1.43 times more likely to develop AD than the carriers of the G allele (95% CI: 1.23-1.61, p < 0.01). The rs2015 locus single-nucleotide polymorphism (SNP) affected the binding efficiency between miR-376a-5p and miR-8061 and the 3'UTR of the SIRT2 gene, and miR-376a-5p and miR-8061 bound to SIRT2 rs2015 A allele to down-regulate the expression of the SIRT2 protein. The rs2241703 SNP affected the binding efficiency between miR-486-3p and the 3'UTR of SIRT2 gene, and miR-486-3p bound to SIRT2 rs2241703 A allele to down-regulate SIRT2 protein expression. The SIRT2 gene rs2015 and rs2241703 loci SNPs are associated with the risk of AD. The rs2015 locus SNP affects regulation of miR-376a-5p and miR-8061 in SIRT2 expression and the rs2241703 locus SNP affects regulation of miR-486-3p in SIRT2, but further studies are needed to verify this mechanism.
本研究旨在探讨位于 Sirtuin 2(SIRT2)基因非翻译区(UTR)3'端单核苷酸多态性(SNP)与阿尔茨海默病(AD)发病风险之间的相关性,并探讨其潜在机制。本研究共纳入 260 例 AD 患者和 260 例健康对照者。采用 Sanger 测序法对所有参与者的 SIRT2 基因 rs2015 和 rs2241703 位点的基因型进行分析。采用实时定量聚合酶链反应(qRT-PCR)分析 microRNAs(miRNAs)和 SIRT2 mRNA 水平。采用 Western blot 分析 SIRT2 蛋白表达水平。采用双荧光素酶报告基因检测和细胞转染实验检测 miRNA 调控 SIRT2 表达的作用。SIRT2 基因 rs2015 位点 A 等位基因携带者发生 AD 的风险较 C 等位基因携带者降低 0.69 倍(95%置信区间:0.59-0.80,p<0.01)。SIRT2 基因 rs2241703 位点 A 等位基因携带者发生 AD 的风险较 G 等位基因携带者增加 1.43 倍(95%置信区间:1.23-1.61,p<0.01)。rs2015 位点 SNP 影响 miR-376a-5p 和 miR-8061 与 SIRT2 基因 3'UTR 的结合效率,miR-376a-5p 和 miR-8061 结合 SIRT2 rs2015 A 等位基因,下调 SIRT2 蛋白表达。rs2241703 SNP 影响 miR-486-3p 与 SIRT2 基因 3'UTR 的结合效率,miR-486-3p 结合 SIRT2 rs2241703 A 等位基因,下调 SIRT2 蛋白表达。SIRT2 基因 rs2015 和 rs2241703 位点 SNP 与 AD 发病风险相关。rs2015 位点 SNP 影响 miR-376a-5p 和 miR-8061 对 SIRT2 表达的调控,rs2241703 位点 SNP 影响 miR-486-3p 对 SIRT2 的调控,但需要进一步研究来验证这一机制。