title: MiR-20b targets AKT3 and modulates vascular endothelial growth factor-mediated changes in diabetic retinopathy.
abstract: Diabetic retinopathy (DR) is the leading cause of new-onset blindness. The roles of microRNAs in diabetic retinopathy are largely unknown. The aim of this study is to investigate the role of miR-20b in DR. Transfection of miR-20b mimic in high glucose (HG)-treated human retinal endothelial cells (HRECs) increased miR-20b expression and decreased the expression level of VEGF mRNA, while transfection of miR-20b inhibitor in control HRECs reduced the miR-20b expression with a corresponding increase of VEGF mRNA. In vitro functional assay showed that transfection of miR-20b mimic prevented HG-induced increase in transendothelial permeability and tube formation in HRECs. Transfection of miR-20b inhibitor or treatment of VEGF increased transendothelial permeability and tube formation in control HRECs. Luciferase reported assay showed that AKT3 is a target of miR-20b. Transfection of miR-20b mimic prevented the up-regulation of AKT3 induced by HG without changing the protein levels of other isoforms of AKT, and silencing of AKT3 caused decrease of VEGF mRNA and protein levels as well as prevented HG-induced increase in transendothelial permeability and tube formation. Finally, we showed that miR-20b was down-regulated in the retina and retinal endothelial cells in diabetic rats, with a correlated up-regulation of VEGF and AKT3. Intravitreal injection of miR-20b mimic in the diabetic rat significantly increased the miR-20b expression and decreased the expression levels of AKT3 and VEGF in the retina tissues, and intravitreal delivery of AKT3 siRNA in the diabetic rat significantly decreased the expressions of AKT3 and VEGF. Collectively, miR-20b is important for the regulation of VEGF-mediated changes in HRECs and rat retinal tissues under hyperglycemic conditions possibly via targeting AKT3.
authors.affiliation: Shenzhen Eye Hospital, Affiliated Shenzhen Eye Hospital of Jinan University, Joint College of Optometry, Shenzhen University, Shenzhen Key Laboratory of Ophthalmology, Shenzhen 518040, China qinbo1024@sina.com.
authors.forename: Bo
authors.initials: B
authors.lastname: Qin
authors.affiliation: Shenzhen Eye Hospital, Affiliated Shenzhen Eye Hospital of Jinan University, Joint College of Optometry, Shenzhen University, Shenzhen Key Laboratory of Ophthalmology, Shenzhen 518040, China.
authors.forename: Jinwen
authors.initials: J
authors.lastname: Liu
authors.affiliation: Shenzhen Eye Hospital, Affiliated Shenzhen Eye Hospital of Jinan University, Joint College of Optometry, Shenzhen University, Shenzhen Key Laboratory of Ophthalmology, Shenzhen 518040, China.
authors.forename: Shenwen
authors.initials: S
authors.lastname: Liu
authors.affiliation: Shenzhen Eye Hospital, Affiliated Shenzhen Eye Hospital of Jinan University, Joint College of Optometry, Shenzhen University, Shenzhen Key Laboratory of Ophthalmology, Shenzhen 518040, China.
authors.forename: Baijun
authors.initials: B
authors.lastname: Li
authors.affiliation: Shenzhen Eye Hospital, Affiliated Shenzhen Eye Hospital of Jinan University, Joint College of Optometry, Shenzhen University, Shenzhen Key Laboratory of Ophthalmology, Shenzhen 518040, China.
authors.forename: Jing
authors.initials: J
authors.lastname: Ren
mesh_terms: D000818:Animals; D002478:Cells, Cultured; D003930:Diabetic Retinopathy / Q000378:metabolism / Q000473:pathology*; D006801:Humans; D051057:Proto-Oncogene Proteins c-akt / Q000378:metabolism*; D012333:RNA, Messenger / Q000235:genetics; D051381:Rats; D012160:Retina / Q000378:metabolism; D015854:Up-Regulation; D042461:Vascular Endothelial Growth Factor A / Q000235:genetics / Q000378:metabolism / Q000502:physiology*
chemical_list: D012333:RNA, Messenger; D042461:Vascular Endothelial Growth Factor A; C491754:AKT3 protein, human; D051057:Proto-Oncogene Proteins c-akt
keywords: AKT3; HRECs; VEGF; diabetic retinopathy; miR-20b; retina