LncRNA KCNQ1OT1 敲低通过 miR-26a-5p/ITGAV/TGF-β/Smad3 轴抑制人晶状体上皮细胞的活力、迁移和上皮-间充质转化。
LncRNA KCNQ1OT1 knockdown inhibits viability, migration and epithelial-mesenchymal transition in human lens epithelial cells via miR-26a-5p/ITGAV/TGF-beta/Smad3 axis.
机构信息
The First Affiliated Hospital of University of Science and Technology of China, Hefei, Anhui, 230001, China.
Department of Ophthalmology, The First Affiliated Hospital of University of Science and Technology of China, Hefei, Anhui, 230001, China.
出版信息
Exp Eye Res. 2020 Nov;200:108251. doi: 10.1016/j.exer.2020.108251. Epub 2020 Sep 17.
BACKGROUND
Long noncoding RNA potassium voltage-gated channel subfamily Q member 1 opposite strand/antisense transcript 1 (KCNQ1OT1) takes part in diabetic cataract progression. This research aims to analyze the function and mechanism of KCNQ1OT1 on viability, migration and epithelial-mesenchymal transition (EMT) in lens epithelial cells.
METHODS
20 diabetic cataract posterior lens capsule tissues and normal samples were collected. Lens epithelial cells (SRA01/04) were stimulated via high glucose (HG). The levels of KCNQ1OT1, miR-26a-5p, integrin αV (ITGAV), TGF-β, Smad3 and phosphorylated (p)-Smad3 were measured via quantitative real-time polymerase chain reaction or Western blot. Cell viability, migration and EMT were analyzed via MTT, wound healing, transwell and Western blot assays. The target relationship between miR-26a-5p and KCNQ1OT1 or ITGAV was determined via luciferase reporter assay.
RESULTS
KCNQ1OT1 was up-regulated and miR-26a-5p level was reduced in diabetic cataract tissues and HG-treated SRA01/04 cells. Silence of KCNQ1OT1 or miR-26a-5p up-regulation repressed cell viability, migration and EMT in SRA01/04 cells stimulated via HG. KCNQ1OT1 could target miR-26a-5p and controlled cell viability, migration and EMT via regulating miR-26a-5p. ITGAV was targeted via miR-26a-5p and positively regulated via KCNQ1OT1. ITGAV overexpression promoted cell viability, migration and EMT in HG-treated SRA01/04 cells, which were mitigated by KCNQ1OT1 silence. KCNQ1OT1 knockdown mitigated HG-induced the activation of TGF-β/Smad3 signaling by regulating miR-26a-5p.
CONCLUSION
KCNQ1OT1 knockdown represses cell viability, migration and EMT through miR-26a-5p/ITGAV/TGF-β/Smad3 axis in SRA01/04 cells under HG condition, providing a new target for the treatment of diabetic cataract.
背景
长链非编码 RNA 钾电压门控通道亚家族 Q 成员 1 反义转录本 1（KCNQ1OT1）参与糖尿病性白内障的进展。本研究旨在分析 KCNQ1OT1 对晶状体上皮细胞活力、迁移和上皮-间充质转化（EMT）的作用和机制。
方法
收集 20 例糖尿病性白内障后晶状体囊组织和正常样本。用高糖（HG）刺激晶状体上皮细胞（SRA01/04）。通过实时定量聚合酶链反应或 Western blot 检测 KCNQ1OT1、miR-26a-5p、整合素 αV（ITGAV）、TGF-β、Smad3 和磷酸化（p）-Smad3 的水平。通过 MTT、划痕愈合、Transwell 和 Western blot 测定细胞活力、迁移和 EMT。通过荧光素酶报告基因测定确定 miR-26a-5p 与 KCNQ1OT1 或 ITGAV 的靶关系。
结果
糖尿病性白内障组织和 HG 处理的 SRA01/04 细胞中 KCNQ1OT1 上调，miR-26a-5p 水平降低。HG 刺激的 SRA01/04 细胞中沉默 KCNQ1OT1 或上调 miR-26a-5p 抑制细胞活力、迁移和 EMT。KCNQ1OT1 可靶向 miR-26a-5p，并通过调节 miR-26a-5p 控制细胞活力、迁移和 EMT。ITGAV 被 miR-26a-5p 靶向并被 KCNQ1OT1 正向调控。ITGAV 过表达促进 HG 处理的 SRA01/04 细胞的细胞活力、迁移和 EMT，而 KCNQ1OT1 沉默减轻了这些作用。KCNQ1OT1 敲低通过调节 miR-26a-5p 减轻 HG 诱导的 TGF-β/Smad3 信号激活。
结论
在 HG 条件下，沉默 KCNQ1OT1 通过 miR-26a-5p/ITGAV/TGF-β/Smad3 轴抑制 SRA01/04 细胞的活力、迁移和 EMT，为糖尿病性白内障的治疗提供了新的靶点。
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