title: Probable Chemical Hypoxia Effects on Progress of CNV Through Induction of Promoter CpG Demethylation and Overexpression of IL17RC in Human RPE Cells.
abstract: To survey the changes of promoter CpG methylation status and mRNA expression of IL17RC (interleukin 17 receptor C) gene in retinal pigment epithelium (RPE) cells under chemical hypoxia condition for choroidal neovascularization (CNV) modeling in vitro.
  
  
  
  RPE cells were cultured in both untreated as a control group and treated by cobalt chloride media as a hypoxia group for various concentrations (100-150μM) and times (24-36 hrs.) To confirm chemical hypoxia condition, mRNA expression of HIF (Hypoxia Inducible Factor) -1α, -2α, and Vascular Endothelial Growth Factor (VEGF) was compared between two groups by Real-time PCR. Also, in normoxia and hypoxia conditions, IL17RC expression changes and promoter CpG methylation status were evaluated by Real-time PCR and methylation-specific PCR (MSP) techniques, respectively.
  
  
  
  Overexpression of HIF-1α, HIF-2α, and VEGF was significant in hypoxia versus normoxia conditions. Our data showed overexpression of IL17RC (2.1- to 6.3-fold) and decreasing of its promoter methylation in comparison with hypoxia and normoxia conditions. It was found that there are significant association between promoter methylation status and expression of IL17RC in chemical hypoxia condition.
  
  
  
  Therefore, methylation of IL17RC could play as a marker in CNV and degeneration of RPE cells in vitro. Additionally, HIF-α and methylation phenomena may be considered as critical targets for blocking in angiogenesis of age-related degeneration in future studies.
authors.affiliation: a Molecular Medicine Department , Institute of Medical Biotechnology, National Institute of Genetic Engineering and Biotechnology (NIGEB) , Tehran , Iran.|b Department of Medical Genetic , Medical Faculty, Tabriz University of Medical Sciences , Tabriz , Iran.
authors.forename: Mohammad Reza
authors.initials: MR
authors.lastname: Alivand
authors.affiliation: a Molecular Medicine Department , Institute of Medical Biotechnology, National Institute of Genetic Engineering and Biotechnology (NIGEB) , Tehran , Iran.
authors.forename: Farzaneh
authors.initials: F
authors.lastname: Sabouni
authors.affiliation: a Molecular Medicine Department , Institute of Medical Biotechnology, National Institute of Genetic Engineering and Biotechnology (NIGEB) , Tehran , Iran.
authors.forename: Zahra-Soheila
authors.initials: ZS
authors.lastname: Soheili
mesh_terms: D002478:Cells, Cultured; D020256:Choroidal Neovascularization / Q000235:genetics* / Q000378:metabolism; D019175:DNA Methylation; D005786:Gene Expression Regulation; D006801:Humans; D000860:Hypoxia / Q000209:etiology / Q000378:metabolism* / Q000473:pathology; D012333:RNA, Messenger / Q000235:genetics*; D060888:Real-Time Polymerase Chain Reaction; D018123:Receptors, Interleukin / Q000096:biosynthesis / Q000235:genetics*; D055213:Retinal Pigment Epithelium / Q000378:metabolism* / Q000473:pathology
chemical_list: C448419:IL17RC protein, human; D012333:RNA, Messenger; D018123:Receptors, Interleukin
keywords: Age-related macular degeneration; DNA methylation; IL17RC; chemical hypoxia; choroidal neovascularization; cobalt chloride; retinal pigment epithelium