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敲低长链非编码 RNA TUG1 通过调控 miR-505-3p/VEGFA 抑制角膜血管生成。

Knockdown of lncRNA TUG1 suppresses corneal angiogenesis through regulating miR-505-3p/VEGFA.

机构信息

Department of Ophthalomology, Tongji Hospital, Tongji Medical College, Huazhong University of Science and Technology, Wuhan 430030, China.

Department of Ophthalomology, Tongji Hospital, Tongji Medical College, Huazhong University of Science and Technology, Wuhan 430030, China.

出版信息

Microvasc Res. 2021 Nov;138:104233. doi: 10.1016/j.mvr.2021.104233. Epub 2021 Aug 17.

Abstract

OBJECTIVES

Vascular endothelial growth factor A (VEGFA) is one of the major factors initiating and regulating angiogenesis. LncRNA taurine up-regulated gene 1 (TUG1) has been implicated in the pathological neovascularization. The aim of this study is to explore the function of TUG1 in regulating VEGFA-mediated angiogenesis in endothelial cells.

METHODS

A total of 12 corneal neovascularization (CRNV) samples were collected form patient undergoing corneal transplantation at Tongji Hospital, Wuhan, China. qRT-PCR and Western blotting were performed to examine gene expression and protein levels. Human umbilical vein endothelial cells (HUVECs) were used as an in vitro angiogenesis model. CCK-8 proliferation assay was used to determine cell proliferation capacity and wound healing was performed to analyze cell migration ability. Dual luciferase reporter assay was used for functional interaction validation between miR-505-3p and its targets. The in vitro angiogenic potential was evaluated by tube formation assay.

RESULTS

TUG1 and VEGFA were upregulated in CRNV tissues and VEGFA-treated HUVECs. TUG1 knockdown inhibited proliferation, migration and tube formation capacity of HUVECs. TUG1 regulated the angiogenesis of HUVECs by modulating VEGFA expression through targeting miR-505-3p.

CONCLUSIONS

Our results suggest that lncRNA TUG1 promotes the angiogenesis of HUVECs through modulating miR-505-3p/VEGFA axis.

摘要

目的

血管内皮生长因子 A(VEGFA)是启动和调节血管生成的主要因素之一。长链非编码 RNA 牛磺酸上调基因 1(TUG1)已被牵涉到病理性新血管形成中。本研究旨在探讨 TUG1 在调节内皮细胞中 VEGFA 介导的血管生成中的作用。

方法

共收集了 12 例来自中国武汉同济医院行角膜移植术患者的角膜新生血管(CRNV)样本。qRT-PCR 和 Western blot 用于检测基因表达和蛋白水平。人脐静脉内皮细胞(HUVEC)被用作体外血管生成模型。CCK-8 增殖实验用于测定细胞增殖能力,划痕实验用于分析细胞迁移能力。双荧光素酶报告实验用于验证 miR-505-3p 与其靶标的功能相互作用。通过管形成实验评估体外血管生成潜能。

结果

TUG1 和 VEGFA 在 CRNV 组织和 VEGFA 处理的 HUVECs 中上调。TUG1 敲低抑制了 HUVECs 的增殖、迁移和管形成能力。TUG1 通过靶向 miR-505-3p 调节 VEGFA 表达来调节 HUVECs 的血管生成。

结论

我们的结果表明,lncRNA TUG1 通过调节 miR-505-3p/VEGFA 轴促进 HUVECs 的血管生成。

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