LncRNA HOTAIR targets miR-126-5p to promote the progression of Parkinson's disease through RAB3IP
LncRNA HOTAIR targets miR-126-5p to promote the progression of Parkinson's disease through RAB3IP
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DOI：
10.1515/hsz-2018-0431
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发表时间：
2019-09-01
影响因子：
3.7
通讯作者：
Lin, Yingjie
中科院分区：
文献类型：
--
作者：
Lin, Qiuyu;Hou, Sen;Lin, Yingjie
帕金森病（Parkinson's disease，PD）是一种以中脑多巴胺能（dopaminergic，DA）神经元变性和死亡为特征的常见神经系统疾病，长链非编码RNA HOTAIR已被证明影响PD的疾病进展。本研究旨在进一步阐明HOTAIR治疗PD的分子机制。利用生物信息学分析确定HOTAIR在PD中的潜在下游靶点。使用荧光素酶测定和RNA结合蛋白免疫沉淀（RIP）测定来验证竞争性内源RNA（ceRNA）之间结合位点的存在。实时定量聚合酶链反应（qRT-PCR）和Western blotting结果显示，在PD细胞和PD小鼠中HOTAIR和RAB 3 IP表达增加，而miR-126- 5 p表达减少。此外，CCK-8测定和流式细胞术分析表明，HOTAIR和RAB 3 IP的敲低以及miR-126- 5 p的过表达显著增加了PD细胞中的细胞增殖并减少了细胞凋亡。此外，体内实验的结果表明，敲低HOTAIR表达增加TH阳性细胞的数量和α-突触核蛋白阳性细胞的数量减少，同时降低DA神经元的凋亡率。我们的研究证实HOTAIR通过以ceRNA依赖性方式调节miR-126- 5 p和RAB 3 IP促进PD进展，并进一步阐明了HOTAIR在PD中的作用机制。
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 alpha-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.