A Proteomics Approach to Investigate miR-153-3p and miR-205-5p Targets in Neuroblastoma Cells.
A Proteomics Approach to Investigate miR-153-3p and miR-205-5p Targets in Neuroblastoma Cells.
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DOI：
10.1371/journal.pone.0143969
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发表时间：
2015
期刊：
影响因子：
3.7
通讯作者：
Møller SG
中科院分区：
文献类型：
--
作者：
Patil KS;Basak I;Pal R;Ho HP;Alves G;Chang EJ;Larsen JP;Møller SG
MicroRNA 是与多种疾病相关的关键调节因子。在 HEK293 细胞中，miR-153-3p 和 miR-205-5p 下调 α-突触核蛋白 (SNCA) 和富含亮氨酸重复激酶 2 (LRRK2)，这两种与帕金森病 (PD) 相关的关键蛋白。我们使用二维凝胶电泳 (2D-PAGE) 与质谱 (MS) 结合来鉴定神经元 SH-SY5Y 细胞中的 miR-153-3p 和 miR-205-5p 靶标谱。我们在 SH-SY5Y 细胞中过度表达并抑制这两种 microRNA，并通过比较蛋白质组学分析，我们从每次分析中定量了约 240 个蛋白质点。结合起来，鉴定出 33 个蛋白质点，显示出显着的丰度变化（p 值 < 0.05）。 miR-153-3p 的调节导致七个蛋白质上调和八个蛋白质下调。 miR-205 调节导致 12 种蛋白上调和 6 种蛋白下调。其中一些蛋白质与神经元过程相关，包括过氧化还原蛋白-2 和 -4、cofilin-1、前折叠蛋白 2、α-烯醇化酶、人核苷二磷酸激酶 B (Nm23) 和 14-3-3 蛋白 epsilon。许多差异表达的蛋白质参与不同的途径，包括代谢、神经营养蛋白信号传导、肌动蛋白细胞骨架调节、HIF-1信号传导和蛋白酶体，表明miR-153-3p和miR-205-5p参与神经母细胞瘤细胞中多种生物过程的调节。
MicroRNAs are key regulators associated with numerous diseases. In HEK293 cells, miR-153-3p and miR-205-5p down-regulate alpha-synuclein (SNCA) and Leucine-rich repeat kinase 2 (LRRK2), two key proteins involved in Parkinson’s disease (PD). We have used two-dimensional gel electrophoresis (2D-PAGE) coupled to mass spectrometry (MS) to identify a spectrum of miR-153-3p and miR-205-5p targets in neuronal SH-SY5Y cells. We overexpressed and inhibited both microRNAs in SH-SY5Y cells and through comparative proteomics profiling we quantified ~240 protein spots from each analysis. Combined, thirty-three protein spots were identified showing significant (p-value < 0.05) changes in abundance. Modulation of miR-153-3p resulted in seven up-regulated proteins and eight down-regulated proteins. miR-205 modulation resulted in twelve up-regulated proteins and six down-regulated proteins. Several of the proteins are associated with neuronal processes, including peroxiredoxin-2 and -4, cofilin-1, prefoldin 2, alpha-enolase, human nucleoside diphosphate kinase B (Nm23) and 14-3-3 protein epsilon. Many of the differentially expressed proteins are involved in diverse pathways including metabolism, neurotrophin signaling, actin cytoskeletal regulation, HIF-1 signaling and the proteasome indicating that miR-153-3p and miR-205-5p are involved in the regulation of a wide variety of biological processes in neuroblastoma cells.