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MicroRNA-29 regulates high-glucose-induced apoptosis in human retinal pigment epithelial cells through PTEN

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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.

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Authors and Affiliations

  1. Department of Ophthalmology, The Second Affiliated Hospital of Chongqing Medical University, 76 Linjiang Road, Chongqing, 400010, China

    Xiaohui Lin, Xiyuan Zhou & Danning Liu

  2. Department of Ophthalmology, The Inner Mongolia Autonomous Region People’s Hospital, 20 Zhaowuda Road, Hohhot, 010010, China

    Lixia Yun, Lina Zhang, Xiaohai Chen, Qinghe Chai & Langen Li

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  1. Xiaohui Lin
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  2. Xiyuan Zhou
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  3. Danning Liu
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  4. Lixia Yun
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  5. Lina Zhang
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  7. Qinghe Chai
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  8. Langen Li
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Correspondence to Xiyuan Zhou.

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Editor: Tetsuji Okamoto

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Lin, X., Zhou, X., Liu, D. et al. MicroRNA-29 regulates high-glucose-induced apoptosis in human retinal pigment epithelial cells through PTEN. In Vitro Cell.Dev.Biol.-Animal 52, 419–426 (2016). https://doi.org/10.1007/s11626-015-9990-z

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  • DOI: https://doi.org/10.1007/s11626-015-9990-z

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