title: MicroRNA-29 regulates high-glucose-induced apoptosis in human retinal pigment epithelial cells through PTEN.
abstract: Hyperglycemia or high-glucose (HG)-induced apoptosis in human retinal pigment epithelial (RPE) cells is a characteristic process in diabetic retinopathy. In our study, we examined whether microRNA-29 (miR-29) may regulate HG-induced RPE cell apoptosis. Human RPE cell line, ARPE-19 cells, was treated with various high concentration of glucose in vitro. HG-induced RPE cell apoptosis was examined by terminal deoxynucleotide transferase-mediated dUTP nick end labeling (TUNEL) assay and miR-29 gene expression by quantitative RT-PCR (qRT-PCR). miR-29 was then downregulated in RPE cells, and its effect on HG-induced apoptosis was examined by TUNEL assay and western blot assay on caspase-7 protein. Association of miR-29 on its downstream target, PTEN, in HG-induced RPE cell apoptosis was evaluated by dual-luciferase assay and qRT-PCR. PTEN was silenced in RPE cells. The effects of PTEN downregulation on miR-29-mediated HG-induced RPE cell apoptosis were also examined by TUNEL and western blot assays. HG induced significant apoptosis in RPE cells in a dose-dependent manner. miR-29 was upregulated by HG in RPE cells. miR-29 downregulation protected HG-induced apoptosis and reduced the production of caspase-7 protein in RPE cells. PTEN was shown to be directly downregulated by HG and then upregulated by miR-29 downregulation in RPE cells. Small interfering RNA (siRNA)-mediated PTEN downregulation reversed the protective effect of miR-29 downregulation on HG-induced RPE cell apoptosis. This study demonstrates that miR-29, through inverse association of PTEN, plays an important role in the process of HG-induced apoptosis in RPE cells.
authors.affiliation: Department of Ophthalmology, The Second Affiliated Hospital of Chongqing Medical University, 76 Linjiang Road, Chongqing, 400010, China.
authors.forename: Xiaohui
authors.initials: X
authors.lastname: Lin
authors.affiliation: Department of Ophthalmology, The Second Affiliated Hospital of Chongqing Medical University, 76 Linjiang Road, Chongqing, 400010, China. xiyuan.zhou@aol.com.
authors.forename: Xiyuan
authors.initials: X
authors.lastname: Zhou
authors.affiliation: Department of Ophthalmology, The Second Affiliated Hospital of Chongqing Medical University, 76 Linjiang Road, Chongqing, 400010, China.
authors.forename: Danning
authors.initials: D
authors.lastname: Liu
authors.affiliation: Department of Ophthalmology, The Inner Mongolia Autonomous Region People's Hospital, 20 Zhaowuda Road, Hohhot, 010010, China.
authors.forename: Lixia
authors.initials: L
authors.lastname: Yun
authors.affiliation: Department of Ophthalmology, The Inner Mongolia Autonomous Region People's Hospital, 20 Zhaowuda Road, Hohhot, 010010, China.
authors.forename: Lina
authors.initials: L
authors.lastname: Zhang
authors.affiliation: Department of Ophthalmology, The Inner Mongolia Autonomous Region People's Hospital, 20 Zhaowuda Road, Hohhot, 010010, China.
authors.forename: Xiaohai
authors.initials: X
authors.lastname: Chen
authors.affiliation: Department of Ophthalmology, The Inner Mongolia Autonomous Region People's Hospital, 20 Zhaowuda Road, Hohhot, 010010, China.
authors.forename: Qinghe
authors.initials: Q
authors.lastname: Chai
authors.affiliation: Department of Ophthalmology, The Inner Mongolia Autonomous Region People's Hospital, 20 Zhaowuda Road, Hohhot, 010010, China.
authors.forename: Langen
authors.initials: L
authors.lastname: Li
mesh_terms: D017209:Apoptosis / Q000187:drug effects*; D001483:Base Sequence; D020169:Caspases / Q000378:metabolism; D002460:Cell Line; D019610:Cytoprotection / Q000187:drug effects; D015536:Down-Regulation / Q000187:drug effects; D004847:Epithelial Cells / Q000187:drug effects / Q000378:metabolism*; D005947:Glucose / Q000494:pharmacology*; D006801:Humans; D035683:MicroRNAs / Q000235:genetics / Q000378:metabolism*; D051059:PTEN Phosphohydrolase / Q000037:antagonists & inhibitors / Q000378:metabolism*; D055213:Retinal Pigment Epithelium / Q000166:cytology*; D015854:Up-Regulation / Q000187:drug effects
chemical_list: C532045:MIRN29a microRNA, human; D035683:MicroRNAs; D051059:PTEN Phosphohydrolase; C494929:PTEN protein, human; D020169:Caspases; D005947:Glucose
keywords: Apoptosis; High glucose; PTEN; Retinal pigment epithelial cells; miR-29
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