Post-transcriptional regulation of cardiac sodium channel gene SCN5A expression and function by miR-192-5p.
Post-transcriptional regulation of cardiac sodium channel gene SCN5A expression and function by miR-192-5p.
复制标题
DOI：
10.1016/j.bbadis.2015.07.016
复制
发表时间：
2015-10
期刊：
影响因子：
--
通讯作者：
Huang Z
中科院分区：
文献类型：
--
作者：
Zhao Y;Huang Y;Li W;Wang Z;Zhan S;Zhou M;Yao Y;Zeng Z;Hou Y;Chen Q;Tu X;Wang QK;Huang Z
SCN 5A基因编码心脏钠通道Nav1.5，突变时可导致致命性室性心律失常/猝死和心房颤动（AF）。microRNAs（miRNAs）是基因表达的重要转录后调节因子，参与多种疾病的发病机制。然而，关于miRNA对SCN 5A的调控知之甚少。在这里，我们揭示了一种新的转录后调控机制，通过miR-192- 5 p的表达和功能的SCN 5A/Nav 1. 5。生物信息学分析显示，人和恒河猴SCN 5A的3′-UTR含有miR-192- 5 p的靶结合位点，而大象、猪、兔、小鼠和大鼠SCN 5A的3 ′-UTR则没有。双荧光素酶报告基因分析表明，该位点对人SCN 5A的下调至关重要。通过Western印迹分析和电生理学研究，我们证明miR-192- 5 p显著降低了SCN 5A和Nav1.5的表达以及Nav1.5产生的峰值钠电流密度INa。值得注意的是，原位杂交、免疫组织化学和实时qPCR分析显示，miR-192- 5 p在AF患者的组织样本中上调，这与SCN 5A/Nav1.5的下调相关。这些结果证明了miR-192- 5 p在Nav1.5的转录后调节中的重要转录后作用，揭示了miR-192- 5 p在心脏生理学和疾病中的新作用，并为针对INa降低的疾病的新型基于miRNA的抗肿瘤治疗提供了新靶点。
The SCN5A gene encodes cardiac sodium channel Nav1.5 and causes lethal ventricular arrhythmias/sudden death and atrial fibrillation (AF) when mutated. MicroRNAs (miRNAs) are important post-transcriptional regulators of gene expression, and involved in the pathogenesis of many diseases. However, little is known about the regulation of SCN5A by miRNAs. Here we reveal a novel post-transcriptional regulatory mechanism for expression and function of SCN5A/Nav1.5 via miR-192-5p. Bioinformatic analysis revealed that the 3′-UTR of human and rhesus SCN5A, but not elephant, pig, rabbit, mouse, and rat SCN5A, contained a target binding site for miR-192-5p and dual luciferase reporter assays showed that the site was critical for down-regulation of human SCN5A. With Western blot assays and electrophysiological studies, we demonstrated that miR-192-5p significantly reduced expression of SCN5A and Nav1.5 as well as peak sodium current density INa generated by Nav1.5. Notably, in situ hybridization, immunohistochemistry and real-time qPCR analyses showed that miR-192-5p was up-regulated in tissue samples from AF patients, which was associated with down-regulation of SCN5A/Nav1.5. These results demonstrate an important post-transcriptional role of miR-192-5p in post-transcriptional regulation of Nav1.5, reveal a novel role of miR-192-5p in cardiac physiology and disease, and provide a new target for novel miRNA-based antiarrhythmic therapy for diseases with reduced INa.