miR-181 interacts with signaling adaptor molecule DENN/MADD and enhances TNF-induced cell death.
miR-181 interacts with signaling adaptor molecule DENN/MADD and enhances TNF-induced cell death.
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DOI：
10.1371/journal.pone.0174368
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发表时间：
2017
期刊：
影响因子：
3.7
通讯作者：
Noorbakhsh F
中科院分区：
文献类型：
--
作者：
Ghorbani S;Talebi F;Ghasemi S;Jahanbazi Jahan Abad A;Vojgani M;Noorbakhsh F
MicroRNA是一类非编码小分子RNA，通过mRNA降解或翻译抑制来调控蛋白质编码转录本的表达。许多报道强调了miRNA在调节细胞死亡途径中的作用，包括参与诱导细胞凋亡的基因的表达。肿瘤坏死因子α（TNF-α）是一种促炎细胞因子，可通过其受体TNFR 1发送促死亡信号。包括DENN/MADD衔接蛋白在内的多种衔接分子已被证明通过将MAP激酶募集至TNFR 1和激活促存活NFκB信号传导来调节TNF-α促死亡信号传导。在此，我们研究了microRNA-181（miR-181）在调节DENN/MADD表达水平中的作用及其对TNF-α诱导的细胞死亡的后续影响。使用生物信息学分析，然后使用荧光素酶报告基因测定，我们表明miR-181与DENN/MADD转录物的3' UTR相互作用。miR-181过表达还导致L929鼠成纤维细胞中内源性DENN/MADD mRNA水平降低。对转染miR-181的细胞的流式细胞术分析显示，该miRNA加重了TNF-α引起的线粒体膜电位损失。这些结果与TNF-α处理后L929细胞凋亡增加有关。总体而言，这些数据表明miR-181在调节TNF-α促死亡信号传导中的潜在作用，这在与组织变性和细胞死亡相关的炎症性疾病的发病机制和治疗方面可能具有重要意义。
MicroRNAs are small noncoding RNAs, which regulate the expression of protein coding transcripts through mRNA degradation or translational inhibition. Numerous reports have highlighted the role of miRNAs in regulating cell death pathways including the expression of genes involved in the induction of apoptosis. Tumor necrosis factor alpha (TNF-α) is a proinflammatory cytokine which can send pro-death signals through its receptor TNFR1. Diverse adaptor molecules including DENN/MADD adaptor protein have been shown to modulate TNF-α pro-death signaling via recruitment of MAP kinases to TNFR1 and activation of pro-survival NFκB signaling. Herein, we investigated the role of microRNA-181 (miR-181) in regulating DENN/MADD expression levels and its subsequent effects on TNF-α-induced cell death. Using bioinformatics analyses followed by luciferase reporter assays we showed that miR-181 interacts with the 3’ UTR of DENN/MADD transcripts. miR-181 overexpression also led to decreased endogenous DENN/MADD mRNA levels in L929 murine fibroblasts. Flow cytometric analysis of miR-181 transfected cells showed this miRNA accentuates mitochondrial membrane potential loss caused by TNF-α. These findings were associated with enhanced apoptosis of L929 cells following TNF-α treatment. Overall, these data point to the potential role of miR-181 in regulating TNF-α pro-death signaling, which could be of importance from pathogenesis and therapeutic perspectives in inflammatory disorders associated with tissue degeneration and cell death.