Unknown,Transcriptomics,Genomics,Proteomics

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Oxidative stress related long non-coding RNAs (lncRNAs) in human umbilical vein endothelial cells

ABSTRACT: Long non-coding RNAs (lncRNAs) exhibit a poor interspecies conservation and often show spatial- and temporal-specific expression patterns. What, if any, role they have in oxidative stress remains unknown. To identify potential roles for lncRNAs, we examined their expression in normal and H2O2-treated human umbilical vein endothelial cells. Oxidative stress related lncRNAs were generated by deep sequencing, using Illumina HiSeq 2000 or 2500 platform. Sequencing of the cDNA libraries from H2O2-treated HUVECs generated 12.5 million uniquely valid reads, meanwhile, 10.2 million valid fragments were obtained from control group in our experiment. A total of 10, 765 known and 30, 629 novel putative lncRNAs were identified according to RNA-Seq. Among them, 2, 091 of known and 25, 800 of novel lncRNAs were differentially expressed in H2O2-treated HUVECs compared with control HUVECs, and 12 of these were validated with qRTâPCR. Taken together, our findings provide evidence differentially expressed lncRNAs were mediated by oxidative stress in HUVECs, it is, therefore, likely that aberrant expression of lncRNAs, at least in part, participate in the process of endothelial injury caused by oxidative stress. Examination of lncRNAs in the oxidative-stressed human umbilical vein endothelial cells

ORGANISM(S): Homo sapiens

SUBMITTER: Rui Zhang

PROVIDER: E-GEOD-78275 | biostudies-arrayexpress |

REPOSITORIES: biostudies-arrayexpress

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Project description:Purpose: Excess oxidative stress (OS) impairs endothelial function and plays an important role in vascular diseases, diabetes, and neuronal disorders. Several consequences of OS including cell recovery and apoptosis have been described previously. In this study, we report systems model of the temporal dynamics of the oxidative stress response in Human Umbilical Vein Endothelial Cells (HUVECs) and characterize HMOX1 as a master regulator in orchestrating the response to oxidative stress. Methods: Following stress induction by hydrogen peroxide (HP), we carried out dose-dependent phenotypic assays and time series measurements of transcripts in HUVECs. To explore the role of HMOX1, HUVECs were transfected with scrambled siRNA or si-HMOX1 and then treated with hydrogen peroxide. Total RNA was collected from Scrambled-untreated, scrambled-H2O2, and si-HMOX1-H2O2 samples for RNAseq. Novel findings were validated by qPCR and functional assays. Results: In this study we have identified three phases of stress response (acute response from 1-2 hours, the transition from 4-6 hours, and the chronic stress response from 8-16 hours). We observe stable cell cycle arrest and impaired DNA replication/repair from 8-16 hours along with and increased resistance to apoptosis from 2-16 hours. Knockdown of HMOX1 prior to HP treatment releases the resistance to apoptosis. Similarly, many cellular processes switch their direction of regulation in HMOX1 deficient cells upon HP treatment (i.e. autophagy, mitochondria, cell surface signaling, expression of histone modifying enzymes and long-noncoding RNAs etc). Conclusions: Endothelial cell fate decisions are critically important for vascular health. In this study we provide a temporal model characterizing the transition from the acute stress response to the chronic response. The nexus of cell function and dysfunction is characterized by the concurrent activation of survival and death pathways. We explore the master regulatory role of HMOX1 and identify novel mechanisms by which HMOX1 may regulate cell fate decisions.

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Oxidative stress related long non-coding RNAs (lncRNAs) in human umbilical vein endothelial cells

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