Article Text
Abstract
Aim To elucidate the role of microRNA-20a-5p (miR-20a-5p) in the pathogenesis of Vogt-Koyanagi-Harada (VKH) disease.
Methods Quantitative real-time PCR was used to quantify miR-20a-5p expression in CD4+ T cells from patients with active VKH and normal controls. The promoter methylation status of miR-20a-5p was detected by bisulfite sequencing PCR. Targets were evaluated by a luciferase reporter assay. The functional effects of miR-20a-5p on CD4+ T cells from patients with active VKH were assessed by upregulation or downregulation of its expression using liposomes.
Results The miR-20a-5p level was significantly decreased in CD4+ T cells from patients with active VKH as compared with normal controls. The two genes, oncostatin M (OSM) and C-C motif chemokine ligand 1 (CCL1), were identified as targets of miR-20a-5p. The upregulation of miR-20a-5p significantly suppressed interleukin 17 (IL-17) production in CD4+ T cells from patients with active VKH, whereas downregulation of miR-20a-5p exhibited an inverse effect. In addition, overexpression of OSM and CCL1 could rescue the effect of the upregulation of miR-20a-5p. Moreover, the level of miR-20a-5p was reduced in response to hypermethylation of the promoter. Further study showed that miR-20a-5p suppressed the activity of the phosphoinositide 3-kinase-AKT pathway.
Conclusions Our findings indicate that downregulation of miR-20a-5p is caused by promoter hypermethylation. MiR-20a-5p could also suppress the production of IL-17 by targeting OSM and CCL1 production in CD4+ T cells in patients with active VKH.
- experimental laboratory
- immunology
- inflammation
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Footnotes
Contributors RC, PY: conceived the idea and designed the experiments. CZ, QC: sample collection. RC, SY, QW: performed all the experiments. GY: flow cytometry test. RC, YH: analysed the data. RC, XT: wrote the manuscript. PY, GS, AK: revised the manuscript.
Funding This work was supported by National Key R&D Program of China (grant no 2016YFC0904000), Natural Science Major International (Regional) Joint Research Project (grant no 81320108009), National Natural Science Foundation Project (grant no 31370893), Chongqing Key Laboratory of Ophthalmology (grant no CSTC, 2008CA5003), National Key Clinical Specialties Construction Program of China, Chongqing Science & Technology Platform and Base Construction Program (grant no cstc2014pt-sy10002) and National Natural Science Foundation Project (grant no 81400390).
Competing interests None declared.
Patient consent Obtained.
Ethics approval The study was approved by the ethical committee of Chongqing Medical University. The tenets of the Declaration of Helsinki were followed during all procedures of the present study.
Provenance and peer review Not commissioned; externally peer reviewed.
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