Document Detail


Modulation of inflammatory markers by miR-146a during replicative senescence in trabecular meshwork cells.
MedLine Citation:
PMID:  20053980     Owner:  NLM     Status:  MEDLINE    
Abstract/OtherAbstract:
PURPOSE: To investigate the alterations in microRNA (miRNA) expression during replicative senescence (RS) in human trabecular meshwork (HTM) cells.
METHODS: Two HTM cell lines were serially passaged until they reached RS. Changes in expression of 30 miRNAs were assessed by real-time quantitative (q)-PCR. The effects of miR-146a on gene expression were analyzed with gene arrays and the results confirmed by real-time q-PCR. Protein levels of IRAK1 and PAI-1 were analyzed by Western blot and those of IL6 and IL8 by ELISA. Senescence-associated markers were monitored by flow cytometry and cell proliferation by BrdU incorporation.
RESULTS: RS of HTM cells was associated with significant changes in expression of 18 miRNAs, including the upregulation of miR-146a. miR-146a downregulated multiple genes associated with inflammation, including IRAK1, IL6, IL8, and PAI-1, inhibited senescence-associated beta-galactosidase (SA-beta-gal) activity and production of intracellular reactive species (iROS), and increased cell proliferation. Overexpression of either IRAK1 or PAI-1 inhibited the effects of miR-146a on cell proliferation and iROS production in senescent cells.
CONCLUSIONS: RS in HTM cells was associated with changes in miRNA expression that could influence the senescent phenotype. Upregulation of the anti-inflammatory miR-146a may serve to restrain excessive production of inflammatory mediators in senescent cells and limit their deleterious effects on the surrounding tissue. Among the different proteins repressed by miR-146a, the inhibition of PAI-1 may act to minimize the effects of senescence on the generation of iROS and growth arrest and prevent alterations of the extracellular proteolytic activity of the TM.
Authors:
Guorong Li; Coralia Luna; Jianming Qiu; David L Epstein; Pedro Gonzalez
Publication Detail:
Type:  Journal Article; Research Support, N.I.H., Extramural     Date:  2010-01-06
Journal Detail:
Title:  Investigative ophthalmology & visual science     Volume:  51     ISSN:  1552-5783     ISO Abbreviation:  Invest. Ophthalmol. Vis. Sci.     Publication Date:  2010 Jun 
Date Detail:
Created Date:  2010-05-20     Completed Date:  2010-06-11     Revised Date:  2011-03-03    
Medline Journal Info:
Nlm Unique ID:  7703701     Medline TA:  Invest Ophthalmol Vis Sci     Country:  United States    
Other Details:
Languages:  eng     Pagination:  2976-85     Citation Subset:  IM    
Affiliation:
Department of Ophthalmology, Duke University, Durham, North Carolina 27710, USA.
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MeSH Terms
Descriptor/Qualifier:
Adult
Blotting, Western
Cell Aging / genetics*
Cell Line
Cell Proliferation
Enzyme-Linked Immunosorbent Assay
Flow Cytometry
Gene Expression Profiling
Genetic Markers
Humans
Interleukin-1 Receptor-Associated Kinases / metabolism
Interleukin-6 / metabolism
Interleukin-8 / metabolism
Membrane Potential, Mitochondrial
MicroRNAs / genetics*
Middle Aged
Oligonucleotide Array Sequence Analysis
Plasminogen Activator Inhibitor 1 / metabolism
Reactive Oxygen Species / metabolism
Reverse Transcriptase Polymerase Chain Reaction
Tissue Donors
Trabecular Meshwork / cytology*,  metabolism*
Transfection
Grant Support
ID/Acronym/Agency:
EY016228/EY/NEI NIH HHS; EY01894/EY/NEI NIH HHS; EY05722/EY/NEI NIH HHS
Chemical
Reg. No./Substance:
0/Genetic Markers; 0/Interleukin-6; 0/Interleukin-8; 0/MIRN146 microRNA, human; 0/MicroRNAs; 0/Plasminogen Activator Inhibitor 1; 0/Reactive Oxygen Species; 0/SERPINE1 protein, human; EC 2.7.1.37/IRAK1 protein, human; EC 2.7.11.1/Interleukin-1 Receptor-Associated Kinases

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