Brainderived neurotrophic factor is a novel target gene of the hsamiR183/96/182 cluster in retinal pigment epithelial cells following visible light exposure
Lightinduced retinal injury is clinically and experimentally welldocumented. It may be categorized into three types: Photothermal, photomechanical and photochemical injuries. To date, the variation in the hsamiR183/96/182 cluster and its potential target genes in human primary retinal pigment epithelial (RPE) cells, following visible light exposure, has not been reported. In the present study, RPE cells were exposed to 4 h of constant visible light. The expression of the hsamiR183/96/182 cluster was determined using reverse transcriptionquantitative polymerase chain reaction (RTqPCR) and its potential target genes were investigated. Additionally, hsamiR183, hsamiR96, hsamiR182 and hasmiR183/96/182 mimics were designed and synthesized in vitro, and transfected into the RPE cells. Subsequently, the expression of brainderived neurotrophic factor (BDNF) mRNA and protein was measured, using RTqPCR and western blotting, respectively. The regulation of miRNAs to the BDNF gene were then validated using a dual luciferase reporter gene assay system. The expression of hsamiR183, hsamiR96 and hsamiR182 significantly increased in RPE cells following 4 h of visible light exposure, compared with RPE cells that had been exposed to dark conditions (P<0.01). Following RPE cell transfection with mimics, BDNF mRNA and protein expression in the RPE cells was significantly downregulated compared with control RPE cells (P<0.05, P<0.01, respectively). Similarly, the ratio of Renilla luciferase/firefly luciferase significantly decreased in the RPE cells of the mimic + wild type (WT) group compared with cells of the psiCHECKTM2 (a vector lacking the sequence of the BDNF gene), wild type and mimic + mutation groups (P<0.05, P<0.01). The present study suggests that BDNF is a target gene of the hasmiR18396182 cluster in RPE cells. The present study suggests an underlying protective mechanism against retinal light injury and may provide a novel target for the prevention and treatment of lightinduced retinal injury.
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Document Type: Research Article
Affiliations: Department of Opthalmology, The Chinese People's Liberation Army General Hospital, Beijing 100000, P.R. China
Publication date: August 1, 2015
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