Research Paper Volume 9, Issue 12 pp 2559—2586
Central role of the p53 pathway in the noncoding-RNA response to oxidative stress
- 1 Molecular Cardiology Laboratory, IRCCS Policlinico S. Donato, 20097, San Donato Milanese, Milan, Italy
- 2 Center for Translational Genomics and BioInformatics, IRCCS San Raffaele Scientific Institute, 20132, Milan, Italy
- 3 Operative Unit of Vascular Surgery I, IRCCS Policlinico S. Donato, 20097, San Donato Milanese, Milan, Italy
- 4 Cardiovascular Research Unit, IRCCS MultiMedica, 20138, Milan, Italy
- 5 Department of Molecular and Traslational Medicine, University of Brescia, 25123, Brescia, Italy
- 6 Vascular Pathology Laboratory, Istituto Dermopatico dell’Immacolata-IRCCS, FLMM, 00167, Rome, Italy
- 7 University of Milan, 20133, Milan, Italy
- 8 Division of Cardiovascular Epigenetics, Department of Cardiology, Goethe University, Frankfurt am Main, 60596, Germany
- 9 Department of Medicine and Surgery, University of Salerno, 84084, Salerno, Italy
Received: October 12, 2017 Accepted: December 1, 2017 Published: December 12, 2017
https://doi.org/10.18632/aging.101341How to Cite
Abstract
Oxidative stress plays a fundamental role in many conditions. Specifically, redox imbalance inhibits endothelial cell (EC) growth, inducing cell death and senescence. We used global transcriptome profiling to investigate the involvement of noncoding-RNAs in these phenotypes. By RNA-sequencing, transcriptome changes were analyzed in human ECs exposed to H2O2, highlighting a pivotal role of p53-signaling. Bioinformatic analysis and validation in p53-silenced ECs, identified several p53-targets among both mRNAs and long noncoding-RNAs (lncRNAs), including MALAT1 and NEAT1. Among microRNAs (miRNAs), miR-192-5p was the most induced by H2O2 treatment, in a p53-dependent manner. Down-modulated mRNA-targets of miR-192-5p were involved in cell cycle, DNA repair and stress response. Accordingly, miR-192-5p overexpression significantly decreased EC proliferation, inducing cell death. A central role of the p53-pathway was also confirmed by the analysis of differential exon usage: Upon H2O2 treatment, the expression of p53-dependent 5’-isoforms of MDM2 and PVT1 increased selectively. The transcriptomic alterations identified in H2O2-treated ECs were also observed in other physiological and pathological conditions where redox control plays a fundamental role, such as ECs undergoing replicative senescence, skeletal muscles of critical limb-ischemia patients and the peripheral-blood mononuclear cells of long-living individuals. Collectively, these findings indicate a prominent role of noncoding-RNAs in oxidative stress response.
Abbreviations
BrdU: bromodeoxyuridine; ChIP-seq: ChIP-sequencing; CLI: critical limb ischemia; EC: endothelial cell; FDR: False Discovery Rate; LLIs: long-living individuals; lncRNAs: long noncoding-RNAs; miRNAs: microRNAs; ncRNAs: noncoding-RNAs; PBMCs: peripheral blood mononuclear cells; SEM: standard error of the mean.