Identification of novel microRNAs in post-transcriptional control of Nrf2 expression and redox homeostasis in neuronal, SH-SY5Y cells.
Identification of novel microRNAs in post-transcriptional control of Nrf2 expression and redox homeostasis in neuronal, SH-SY5Y cells.
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DOI：
10.1371/journal.pone.0051111
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发表时间：
2012
期刊：
影响因子：
3.7
通讯作者：
Mahimainathan L
中科院分区：
文献类型：
--
作者：
Narasimhan M;Patel D;Vedpathak D;Rathinam M;Henderson G;Mahimainathan L
核因子-红细胞2相关因子2 （Nrf2/NFE2L2）是一种氧化还原敏感转录因子，在细胞适应应激过程中起关键作用，并通过启动抗氧化和解毒基因的转录提供细胞防御。虽然蛋白质可以在多个水平上进行调控，但Keap1在翻译后调控点上对Nrf2的控制进行了大量研究。重要的是，基于Nrf2转录后/翻译的调控尚不清楚，迄今为止还没有关于神经元系统中这种机制的报道。在这种背景下，涉及microRNAs （miRs）作用的研究已经到位，microRNAs通常被认为是通过翻译抑制和/或转录后改变对蛋白质产生进行微调的调节剂。在目前的研究中，基于芯片分析、免疫印迹和实时分析，我们首次发现并验证了人类NFE2L2可以被miR153/miR27a/miR142-5p/miR144靶向于神经元、SH-SY5Y细胞。将单个miR模拟物与人Nrf2的WT 3′UTR或Nrf2 3′UTR内靶向种子序列的突变miRNA共转染研究表明，Nrf2是这些miRs的直接调控靶点。此外，miR153/miR27a/miR142-5p/miR144的异位表达以不依赖于Keap1的方式影响Nrf2 mRNA丰度和Nrf2核质浓度，导致Nrf2的反激活能力低下。此外，miRs的强制表达降低了GCLC和GSR的表达，导致Nrf2依赖性氧化还原稳态的改变。最后，基于生物信息学的mirna -疾病网络分析（MDN）以及Nrf2相关病理过程的扩展计算网络分析表明，如果在特定的细胞情况下，这些miR153/miR27a/miR142-5p/miR144中的任何一个单独或作为一个群体发生改变，它都可能影响Nrf2，从而触发和/或决定广泛疾病结局的命运。
Nuclear factor-erythroid 2-related factor 2 (Nrf2/NFE2L2), a redox-sensitive transcription factor plays a critical role in adaptation to cellular stress and affords cellular defense by initiating transcription of antioxidative and detoxification genes. While a protein can be regulated at multiple levels, control of Nrf2 has been largely studied at post-translational regulation points by Keap1. Importantly, post-transcriptional/translational based regulation of Nrf2 is less understood and to date there are no reports on such mechanisms in neuronal systems. In this context, studies involving the role of microRNAs (miRs) which are normally considered as fine tuning regulators of protein production through translation repression and/or post-transcriptional alterations, are in place. In the current study, based on in-silico analysis followed by immunoblotting and real time analysis, we have identified and validated for the first time that human NFE2L2 could be targeted by miR153/miR27a/miR142-5p/miR144 in neuronal, SH-SY5Y cells. Co-transfection studies with individual miR mimics along with either WT 3′ UTR of human Nrf2 or mutated miRNA targeting seed sequence within Nrf2 3′ UTR, demonstrated that Nrf2 is a direct regulatory target of these miRs. In addition, ectopic expression of miR153/miR27a/miR142-5p/miR144 affected Nrf2 mRNA abundance and nucleo-cytoplasmic concentration of Nrf2 in a Keap1 independent manner resulting in inefficient transactivating ability of Nrf2. Furthermore, forced expression of miRs diminished GCLC and GSR expression resulting in alteration of Nrf2 dependent redox homeostasis. Finally, bioinformatics based miRNA-disease network analysis (MDN) along with extended computational network analysis of Nrf2 associated pathologic processes suggests that if in a particular cellular scenario where any of these miR153/miR27a/miR142-5p/miR144 either individually or as a group is altered, it could affect Nrf2 thus triggering and/or determining the fate of wide range of disease outcomes.
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