10 June 2019 : Laboratory Research
microRNA-378a Regulates the Reactive Oxygen Species (ROS)/Phosphatidylinositol 3-Kinases (PI3K)/AKT Signaling Pathway in Human Lens Epithelial Cells and CataractYan Liu1BC, Huanhuan Li1EF, Yao Liu1ADG*
Med Sci Monit 2019; 25:4314-4321
BACKGROUND: Cataract is associated with increased apoptosis of the epithelial cells of the ocular lens. Previous studies have shown that microRNA-378a (miR-378a) has a role in the development of cataract, but the molecular mechanisms remain unclear. This study aimed to investigate the effects of miR-378a in human lens epithelial cells (HLECs) in vitro and normal lens tissues and cataract tissues.
MATERIAL AND METHODS: HLECs were grown in culture. Quantitative reverse transcription polymerase chain reaction (RT-qPCR) and Western blot were used to examine gene expression levels. The MTT and TUNEL assay measured cell growth and apoptosis. Changes in the fluorescence ratio of ethidium to dihydroethidium (E: DHE) and in 6-carboxy-2’,7’-dichlorodihydrofluorescein diacetate (C-H₂DCFDA) were used to detect superoxide (O₂⁻) and hydrogen peroxide (H₂O₂). The expression levels of miR-378a and the superoxide dismutase 1 gene (SOD1) were measured in normal human lens tissues and cataract tissues.
RESULTS: Upregulation of miR-378a reduced the expression of SOD1. Levels of O₂⁻ were upregulated and H₂O₂ was slightly down-regulated by miR-378a. The use of a miR-378a mimic suppressed cell growth and enhanced apoptosis of HLECs, which were reversed by the use of a miR-378a inhibitor. SOD1 overexpression rescued the miR-378a-induced phenotypes of HLEC cells. Treatment with the PI3K inhibitor, LY294002, reversed miR-378a and ROS-regulated proliferation and apoptosis of HLEC cells. Also, miR-378a was upregulated, and SOD1 was down-regulated in human cataract tissues.
CONCLUSIONS: In HLECs, expression of miR-378a regulated ROS and PI3K/AKT signaling, and miR-378a was upregulated, and SOD1 was down-regulated in human cataract tissue.
Keywords: Apoptosis, Cataract, MicroRNAs, Phosphatidylinositol 3-Kinases, Cell Proliferation, Cell Survival, Cells, Cultured, Chromones, Epithelial Cells, Hydrogen Peroxide, Lens, Crystalline, Morpholines, Oxidative Stress, Protein Kinase Inhibitors, Proto-Oncogene Proteins c-akt, Reactive Oxygen Species, Signal Transduction, superoxide dismutase-1, Up-Regulation
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