An Intronic miRNA Regulates Expression of the Human Endothelial Nitric Oxide Synthase Gene and Proliferation of Endothelial Cells by a Mechanism Related to the Transcription Factor SP-1
Open Access
- 5 August 2013
- journal article
- research article
- Published by Public Library of Science (PLoS) in PLOS ONE
- Vol. 8 (8), e70658
- https://doi.org/10.1371/journal.pone.0070658
Abstract
This study was to investigate the molecular mechanisms underlying the 27nt-miRNA-mediated regulation of expression of the endothelial nitric oxide synthase (eNOS) gene. Cell lines overexpressing 27nt-miRNA or its mutant were established by transfecting the miRNA expression vector into the endothelial cells. eNOS mRNA and protein expression were examined by RT-PCR and Western Blotting, respectively. Luciferase activity reporter system was used to study the target of 27nt-miRNA. The results showed that overexpression of 27nt-miRNA significantly inhibited eNOS mRNA level and protein expression, and reduced the eNOS transcriptional efficiency. Such inhibitory effects of 27nt-miRNA were attenuated by the sequence mutations in 27nt-miRNA. Interestingly, the transcription factor SP-1 expression was reduced by 27nt-miRNA. Meanwhile, overxpression of SP-1 protein partially restored eNOS expression, and rescued the 27nt-miRNA-mediated reduction of endothelial cell proliferation. Moreover, certain sites in the SP-1 mRNA were found to be the direct target of 27nt-miRNA by a luciferase reporter system. These results demonstrate that the 27nt-miRNA suppresses eNOS gene expression and SP-1 expression in vascular endothelial cells. The 27nt-miRNA directly target to SP-1 mRNA, thereby contributing to proliferation of endothelial cells.Keywords
This publication has 43 references indexed in Scilit:
- Identification of a two-layer regulatory network of proliferation-related microRNAs in hepatoma cellsNucleic Acids Research, 2012
- Trim71 cooperates with microRNAs to repress Cdkn1a expression and promote embryonic stem cell proliferationNature Communications, 2012
- Vascular Inflammation and Repair: Implications for Re-Endothelialization, Restenosis, and Stent ThrombosisКАРДИОЛОГИЯ УЗБЕКИСТАНА, 2011
- Intronic microRNA suppresses endothelial nitric oxide synthase expression and endothelial cell proliferation via inhibition of STAT3 signalingMolecular and Cellular Biochemistry, 2011
- Endothelial nitric oxide synthase transgenic models of endothelial dysfunctionPflügers Archiv - European Journal of Physiology, 2010
- MicroRNA regulation of a cancer network: Consequences of the feedback loops involving miR-17-92, E2F, and MycProceedings of the National Academy of Sciences of the United States of America, 2008
- Twist-1 regulates the miR-199a/214 cluster during developmentNucleic Acids Research, 2008
- An intronic microRNA silences genes that are functionally antagonistic to its host geneNucleic Acids Research, 2008
- Multilevel Regulation of Gene Expression by MicroRNAsScience, 2008
- Inter- and intra-combinatorial regulation by transcription factors and microRNAsBMC Genomics, 2007