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.

中文翻译：

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LncRNA KCNQ1OT1敲低通过miR-26a-5p / ITGAV / TGF-beta / Smad3轴抑制人晶状体上皮细胞的活力，迁移和上皮间质转化。

背景

与链/反义转录物1（KCNQ1OT1）相反的长非编码RNA钾电压门控通道亚家族Q成员1参与糖尿病性白内障的进展。这项研究旨在分析KCNQ1OT1对晶状体上皮细胞活力，迁移和上皮-间质转化（EMT）的功能和机制。

方法

收集20例糖尿病性白内障后方晶状体囊组织和正常样品。晶状体上皮细胞（SRA01 / 04）通过高葡萄糖（HG）刺激。通过实时定量聚合酶链反应或蛋白质印迹法测量KCNQ1OT1，miR-26a-5p，整联蛋白αV（ITGAV），TGF-β，Smad3和磷酸化（p）-Smad3的水平。通过MTT，伤口愈合，transwell和Western印迹分析来分析细胞活力，迁移和EMT。miR-26a-5p与KCNQ1OT1或ITGAV之间的靶标关系通过荧光素酶报告基因测定法确定。

结果

在糖尿病性白内障组织和经HG处理的SRA01 / 04细胞中，KCNQ1OT1上调，miR-26a-5p水平降低。在通过HG刺激的SRA01 / 04细胞中，KCNQ1OT1或miR-26a-5p上调的沉默抑制了细胞活力，迁移和EMT。KCNQ1OT1可以靶向miR-26a-5p，并通过调节miR-26a-5p来控制细胞活力，迁移和EMT。ITGAV通过miR-26a-5p靶向，并通过KCNQ1OT1受到积极调控。ITGAV的过表达促进了HG处理的SRA01 / 04细胞中的细胞活力，迁移和EMT，这被KCNQ1OT1沉默所缓解。KCNQ1OT1敲低通过调节miR-26a-5p减轻了HG诱导的TGF-β/ Smad3信号传导的激活。

结论

在HG条件下，KCNQ1OT1敲低通过HG条件下的SRA01 / 04细胞中的miR-26a-5p / ITGAV /TGF-β/ Smad3轴抑制细胞活力，迁移和EMT，为糖尿病性白内障的治疗提供了新的靶点。