title: Inhibitory effect of microRNA-34a on retinal pigment epithelial cell proliferation and migration.
abstract: OBJECTIVE
  Retinal pigment epithelial (RPE) cells play important roles in ophthalmologic diseases such as proliferative vitreoretinopathy, AMD, and diabetic retinopathy. MicroRNA-34a (miR-34a) has been reported to be important in the regulation of cell proliferation, migration, differentiation, and apoptosis. In this study, we explored the effects of miR-34a on RPE cells.
  
  
  METHODS
  The expression level of miR-34a in subconfluent and postconfluent ARPE-19 cells was investigated with quantitative real-time PCR. MicroRNA mimic and small interfering RNA (siRNA) were transiently transfected into RPE cells. Transfected RPE cells were analyzed with WST-1 proliferation assay, and their migration was analyzed with transwell assay and in vitro scratch study. The expression or activation of target proteins was detected by Western blotting.
  
  
  RESULTS
  MicroRNA-34a was significantly downregulated in subconfluent ARPE-19 cells compared with postconfluent cells. Introduction of miR-34a inhibited the proliferation and migratory ability of RPE cells without obvious cell apoptosis. In miR-34a transfected cells, many important proliferation and/or migration related molecules such as c-Met, CDK2, CDK4, CDK6, E2F1, and phosphorylated-Cdc2 (p-Cdc2) were downregulated. Small interfering RNA designed to target c-Met also inhibited the proliferation and migration of RPE cells and downregulated CDK2, CDK6, E2F1, and p-Cdc2.
  
  
  CONCLUSIONS
  MicroRNA-34a is downregulated in subconfluent RPE cells. MicroRNA-34a can inhibit the proliferation and migration of RPE cells through downregulation of its targets c-Met and other cell cycle-related molecules. Our results indicated that miR-34a is involved in the regulation of RPE cells.
authors: School of Ophthalmology and Optometry, Eye Hospital, Wenzhou Medical University, Wenzhou, Zhejiang, China.
authors: Qiang
authors: Q
authors: Hou
authors: Jiang
authors: J
authors: Tang
authors: Zhenlian
authors: Z
authors: Wang
authors: Chao
authors: C
authors: Wang
authors: Xiaogang
authors: X
authors: Chen
authors: Ling
authors: L
authors: Hou
authors: Xiang Da
authors: XD
authors: Dong
authors: Lili
authors: L
authors: Tu
mesh_terms: D017209:Apoptosis / Q000235:genetics*; D015153:Blotting, Western; D002453:Cell Cycle; D002465:Cell Movement; D049109:Cell Proliferation; D004247:DNA / Q000235:genetics*; D015536:Down-Regulation; D006801:Humans; D035683:MicroRNAs / Q000096:biosynthesis / Q000235:genetics*; D060888:Real-Time Polymerase Chain Reaction; D055213:Retinal Pigment Epithelium / Q000378:metabolism* / Q000473:pathology; D018630:Vitreoretinopathy, Proliferative / Q000235:genetics* / Q000378:metabolism / Q000473:pathology
chemical_list: C522425:MIRN34 microRNA, human; D035683:MicroRNAs; D004247:DNA
keywords: RPE; c-Met; miR-34a; migration; proliferation