title: MicroRNA-483-3p Inhibits Extracellular Matrix Production by Targeting Smad4 in Human Trabecular Meshwork Cells.
abstract: OBJECTIVE
  This study investigated the effects of microRNA-483-3p (miR-483-3p) on extracellular matrix (ECM) production, and clarified the regulatory mechanism of microRNA-483-3p in human trabecular meshwork cells (HTMCs) under oxidative stress.
  
  
  METHODS
  The expression levels of ECM (fibronectin, laminin, collagen I) in HTMCs under oxidative stress were measured by Western blot. Changes of miR-483-3p expression in HTMCs were evaluated by quantitative polymerase chain reaction (qPCR). After using lentivirus stably expressing pri-miR-483, the effects of miR-483-3p on the ECM were assessed by qPCR and Western blot. Smad4, the potential target of miR-483-3p according to mRNA target-predicting algorithms, was confirmed by luciferase assay and Western blot. Furthermore, the effects of Smad4 knockdown on ECM expression were investigated by qPCR and Western blot.
  
  
  RESULTS
  The mRNA and protein levels of ECM (fibronectin, laminin, collagen I) were upregulated in HTMCs induced by oxidative stress. The expression level of miR-483-3p decreased in HTMCs under oxidative stress, and the ectopic expression of miR-483-3p decreased the levels of ECM. In addition, miR-483-3p targeted Smad4 through two binding sites, resulting in a decrease of Smad4 expression. Furthermore, knockdown of Smad4 reduced the levels of ECM in HTMCs.
  
  
  CONCLUSIONS
  MicroRNA-483-3p has an inhibitory effect on ECM production in HTMCs through downregulating Smad4, which indicates that miR-483-3p may serve as a potential therapeutic target in glaucoma.
authors.affiliation: Eye Hospital Eye Institute, Tianjin Medical University, Tianjin, China 2Department of Cell Biology, 2011 Collaborative Innovation Center of Tianjin for Medical Epigenetics, Laboratory of Epigenetics in Development and Tumorigenesis, Tianjin Research Cente.
authors.forename: Wencui
authors.initials: W
authors.lastname: Shen
authors.affiliation: Department of Cell Biology, 2011 Collaborative Innovation Center of Tianjin for Medical Epigenetics, Laboratory of Epigenetics in Development and Tumorigenesis, Tianjin Research Center of Basic Medical Sciences, Tianjin Key Laboratory of Medical Epigeneti.
authors.forename: Yating
authors.initials: Y
authors.lastname: Han
authors.affiliation: Department of Pathology, Institute of Hematology and Blood Diseases Hospital, Chinese Academy of Medical Sciences and Peking Union Medical College, Tianjin, China.
authors.forename: Bingqing
authors.initials: B
authors.lastname: Huang
authors.affiliation: Eye Hospital Eye Institute, Tianjin Medical University, Tianjin, China 2Department of Cell Biology, 2011 Collaborative Innovation Center of Tianjin for Medical Epigenetics, Laboratory of Epigenetics in Development and Tumorigenesis, Tianjin Research Cente.
authors.forename: Yan
authors.initials: Y
authors.lastname: Qi
authors.affiliation: Eye Hospital Eye Institute, Tianjin Medical University, Tianjin, China 2Department of Cell Biology, 2011 Collaborative Innovation Center of Tianjin for Medical Epigenetics, Laboratory of Epigenetics in Development and Tumorigenesis, Tianjin Research Cente.
authors.forename: Linqi
authors.initials: L
authors.lastname: Xu
authors.affiliation: Eye Hospital Eye Institute, Tianjin Medical University, Tianjin, China.
authors.forename: Ruru
authors.initials: R
authors.lastname: Guo
authors.affiliation: Department of Cell Biology, 2011 Collaborative Innovation Center of Tianjin for Medical Epigenetics, Laboratory of Epigenetics in Development and Tumorigenesis, Tianjin Research Center of Basic Medical Sciences, Tianjin Key Laboratory of Medical Epigeneti.
authors.forename: Xi
authors.initials: X
authors.lastname: Wang
authors.affiliation: Eye Hospital Eye Institute, Tianjin Medical University, Tianjin, China.
authors.forename: Jiantao
authors.initials: J
authors.lastname: Wang
mesh_terms: D015153:Blotting, Western; D049109:Cell Proliferation; D005109:Extracellular Matrix / Q000378:metabolism*; D005786:Gene Expression Regulation; D005901:Glaucoma / Q000235:genetics / Q000378:metabolism / Q000473:pathology; D006801:Humans; D035683:MicroRNAs / Q000096:biosynthesis / Q000235:genetics*; D018384:Oxidative Stress / Q000235:genetics*; D016133:Polymerase Chain Reaction; D012333:RNA, Messenger / Q000235:genetics*; D051901:Smad4 Protein / Q000096:biosynthesis / Q000235:genetics*; D014129:Trabecular Meshwork / Q000378:metabolism* / Q000473:pathology
chemical_list: C548912:MIRN483 microRNA, human; D035683:MicroRNAs; D012333:RNA, Messenger; C497796:SMAD4 protein, human; D051901:Smad4 Protein