Global MicroRNA Expression Profiling Reveals Differential Expression of Target Genes in 6-Hydroxydopamine-injured MN9D Cells
Global MicroRNA Expression Profiling Reveals Differential Expression of Target Genes in 6-Hydroxydopamine-injured MN9D Cells
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DOI：
10.1007/s12017-013-8244-z
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发表时间：
2013-09-01
影响因子：
3.5
通讯作者：
Gao, Dian-Shuai
中科院分区：
文献类型：
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作者：
Li, Li;Chen, Hui-Zhen;Gao, Dian-Shuai
最近的证据表明 microRNA (miRNA) 在神经退行性疾病中发挥着关键作用。然而，人们对这些小 RNA 如何促进多巴胺能神经元凋亡知之甚少。在这里，我们通过 miRCURY (TM) LNA microRNA 阵列分析了经过和未经 6-羟基多巴胺 (6-OHDA) 处理的 MN9D 细胞中 miRNA 的表达。我们确定了 6 个显着降低的 miRNA（miR-668-3p、let-7d-3p、miR-3077-3p、miR-665-5p、miR-99b-3p 和 miR-323-3p），以及 5 个 miRNA（miR-875、miR-207、miR-425-5p、miR-19b-3p 和 miR-19b-3p）。 miR-338-3p）在 6-OHDA 处理后显着升高。其中，有五种已被证明与神经退行性疾病有关。与我们的预测一致，失调的 miRNA 的靶 mRNA，例如过氧化还原蛋白 III (Prx III) 和 Myc，也显示出表达水平的变化。此外，使用双荧光素酶报告基因测定，我们证实 Prx III 是 miR-875 的直接靶基因。总而言之，这些发现表明 6-OHDA 处理后 miRNA 表达发生变化，并表明 miRNA 及其预测靶标在 MN9D 细胞凋亡中具有潜在作用。
Recent evidence indicates that microRNAs (miRNAs) play a key role in neurodegenerative diseases. However, little is known about how these small RNAs contribute to dopaminergic neuronal apoptosis. Here, we profiled the expression of miRNAs in MN9D cells with and without 6-hydroxydopamine (6-OHDA) treatment by miRCURY (TM) LNA microRNA arrays. We identified six miRNAs (miR-668-3p, let-7d-3p, miR-3077-3p, miR-665-5p, miR-99b-3p, and miR-323-3p) that were significantly lower and five miRNAs (miR-875, miR-207, miR-425-5p, miR-19b-3p, and miR-338-3p) that were significantly higher after 6-OHDA treatment. Among them, five have been demonstrated to be implicated in neurodegenerative diseases. Consistent with our prediction, the deregulated miRNA's target mRNAs, such as peroxiredoxin III (Prx III) and Myc, also showed changes in their expression levels. Furthermore, using a dual-luciferase reporter assay, we confirmed that Prx III was a direct target gene of miR-875. Taken together, these findings demonstrate that changes in miRNA expression occur after 6-OHDA treatment and suggest that miRNAs and their predicted targets have a potential role in apoptosis of MN9D cells.