Department of Organic Chemistry, College of Chemistry, Jilin University, No. 2699 Qianjin Street, Changchun 130012, Jilin, China.
Department of Nuclear Medicine, The First Hospital of Jilin University, Changchun 130021, Jilin, China.
Biol Chem. 2019 Aug 27;400(9):1217-1228. doi: 10.1515/hsz-2018-0431.
Parkinson's disease (PD) is a common neurological disorder characterized by dopaminergic (DA) neuron degeneration and death in the midbrain, and the long noncoding RNA HOTAIR has been shown to affect disease progression in PD. In this study, we aimed to further illustrate the molecular mechanism of HOTAIR in PD. Bioinformatics analysis was utilized to determine the potential downstream targets of HOTAIR in PD. Luciferase assay and the RNA Binding Protein Immunoprecipitation (RIP) assay were used to validate the existence of binding sites between competing endogenous RNAs (ceRNAs). Real-time quantitative polymerase chain reaction (qRT-PCR) and Western blotting indicated that HOTAIR and RAB3IP increased while miR-126-5p decreased in PD cells and PD mice. Additionally, the CCK-8 assay and flow cytometric analysis indicated that the knockdown of HOTAIR and RAB3IP and the overexpression of miR-126-5p significantly increased cell proliferation and reduced apoptosis in PD cells. Furthermore, the results of in vivo experiments suggested that knockdown of HOTAIR expression increased the number of TH-positive cells and the number of α-synuclein-positive cells decreased while reducing the apoptosis rate among DA neurons. Our study confirmed that HOTAIR promotes PD progression by regulating miR-126-5p and RAB3IP in a ceRNA-dependent manner and further clarified how HOTAIR works in PD.
帕金森病（PD）是一种常见的神经退行性疾病，其特征是中脑多巴胺能（DA）神经元变性和死亡，长链非编码 RNA HOTAIR 已被证明会影响 PD 的疾病进展。在这项研究中，我们旨在进一步阐明 HOTAIR 在 PD 中的分子机制。我们利用生物信息学分析来确定 HOTAIR 在 PD 中的潜在下游靶标。通过荧光素酶报告基因检测和 RNA 结合蛋白免疫沉淀（RIP）实验来验证竞争内源性 RNA（ceRNA）之间的结合位点是否存在。实时定量聚合酶链反应（qRT-PCR）和 Western blot 表明，在 PD 细胞和 PD 小鼠中，HOTAIR 和 RAB3IP 增加，而 miR-126-5p 减少。此外，CCK-8 检测和流式细胞术分析表明，HOTAIR 和 RAB3IP 的敲低以及 miR-126-5p 的过表达可显著增加 PD 细胞的增殖并减少细胞凋亡。此外，体内实验结果表明，下调 HOTAIR 的表达可增加 TH 阳性细胞的数量，减少 α-突触核蛋白阳性细胞的数量，并降低 DA 神经元的凋亡率。我们的研究证实，HOTAIR 通过 ceRNA 依赖性方式调节 miR-126-5p 和 RAB3IP 促进 PD 进展，并进一步阐明了 HOTAIR 在 PD 中的作用机制。
