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JongHwa Jun; MicroRNA-124-3p Represses Retinal Pigment Epithelial Cells Proliferation and Migration by Targeting RhoG. Invest. Ophthalmol. Vis. Sci. 2016;57(12):3195.
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© ARVO (1962-2015); The Authors (2016-present)
Proliferation and migration of retinal pigment epithelial cells (RPE) are implicated in various retinal pathologies such as epiretinal membrane, AMD, tractional membrane formation in diabetic retinopathy, and proliferative vitreoretinopathy. Small GTPase Rho regulates proliferation or migration, RhoG was reported as a regulatory target of microRNA (miR)-124-3p.
For quantitative measurement of cell viability and BrdU incorporation after transfection of miR-124-3p, cell counting kit-8 and 5-bromo-2'-deoxyuridine (BrdU) enzyme-linked immunosorbent assay were performed, respectively. Ki-67 localizations rate per each nucleus staining were counted after ectopic overexpression of miR-124-3p. Wound healing ability and transwell migration in presence of miR-124-3p mimic or miR-124-3p inhibitor was evaluated. Cytoskeletal alterations were observed by filamentous-actin staining by phalloidin. Cell viability, wound healing, and migration were evaluated after RhoG silencing by small interfering RNA.
MiR-124-3p overexpression distinctively decreased RPE cell viability/BrdU incorporation and Ki-67 localization ratio. Wound healing and migration was impeded by miR-124-3p introduction but miR-124-3p inhibitor promoted these processes. In miR-124-3p transfected retinal cells showed dramatic vanishment of lamellipodium on f-actin staining. Target validation assay showed miR-124-3p transfection decreased RhoG 3’ untranslated region luciferase activity. RhoG siRNA transfection reproduced similar biological change such as decreased cell viability/wound healing/migration in RPE cells.
Down-regulation of RhoG by overexpression of miR-124-3p in RPE cells impeded cell viability/proliferation, wound healing, and migration. In our study, miR-124-3p is involved in regulation of physiology in RPE cells.
This is an abstract that was submitted for the 2016 ARVO Annual Meeting, held in Seattle, Wash., May 1-5, 2016.
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