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下调长链非编码RNA SNHG1通过调控miR-195-5p/血管内皮生长因子A抑制角膜血管生成。

Knockdown of LncRNA SNHG1 Suppresses Corneal Angiogenesis by the Regulation of miR-195-5p/VEGF-A.

作者信息

Hu Xiaofeng, Xing Hongru, Wang Xiaofeng, Du Lei, Huang Yihong, Hao Youguo, Shen Zhibin

机构信息

Department of Ophthalmology, Beijing Chaoyang Hospital, Capital Medical University, Beijing 100020, China.

Neuroelectrophysiology Department, People's Hospital of Shanxi Province, No. 29 Shuangtasi Street, Taiyuan 030000, Shanxi, China.

出版信息

J Ophthalmol. 2021 Oct 19;2021:6646512. doi: 10.1155/2021/6646512. eCollection 2021.

Abstract

LncRNA SNHG1 (SNHG1) has been widely studied as the causative factor of angiogenesis and proliferative agent in gastric, lung, cervical, and hepatocellular carcinomas. However, its significance of angiogenesis and progression of corneal neovascularization (CRNV) is least understood. This study focuses on the molecular mechanisms followed by SNHG1 to establish CRNV and its angiogenesis. Bioinformatics analysis to identify potential miRNA targets of SNHG1 and vascular endothelial growth factor A (VEGF-A) was conducted using StarBase and was subsequently confirmed by the luciferase reporter assay. Relative quantitative expression of SNHG1 in human umbilical vein endothelial cells (HUVECs) was detected through qRT-PCR and western blot analysis. Cell proliferation was detected through CCK-8 assay, whereas migratory abilities of the cells were determined with transwell assay. A capillary-like tube formation assay was performed to detect the tube formation ability of the cells. Following this, relative expression of miR-195-5p and VEGF-A was determined through qRT-PCR and western blot analysis. Results from the experiments manifested upregulated levels of SNHG1 and VEGF-A in HUVECs and CRNV tissues as compared with the control group, whereas downregulated levels of miR-195-5p were measured in the CRNV tissues and HUVECs, suggesting the negative correlation between lncRNA and miRNA. Overexpressed vascular endothelial growth factor promoted cell proliferation and tube formation; however, its silencing leads to inhibition in angiogenesis and proliferation. Potential binding sites of SNHG1 showed miR-195-5p as its direct target and SNHG1 as a sponge for this miRNA. Knockdown and downregulated levels of SNHG1 showed a notable decrease and inhibition in angiogenesis and migration of CRNV cells. The study showed that SNHG1 inhibition significantly reduced cell proliferation, migration, and tube formation in HUVECs transfect with lncRNA SNHG1. Mechanistic insights into the SNHG1 showed that SNHG1 acts as a sponge for miR-195-5p and upregulates the levels of VEGF-A.

摘要

长链非编码RNA SNHG1(SNHG1)已被广泛研究,被认为是胃癌、肺癌、宫颈癌和肝细胞癌血管生成的致病因素及增殖因子。然而,其在角膜新生血管化(CRNV)血管生成和进展中的意义却鲜为人知。本研究聚焦于SNHG1建立CRNV及其血管生成所遵循的分子机制。利用StarBase对SNHG1和血管内皮生长因子A(VEGF - A)的潜在miRNA靶点进行生物信息学分析,随后通过荧光素酶报告基因检测进行验证。通过qRT - PCR和蛋白质印迹分析检测人脐静脉内皮细胞(HUVECs)中SNHG1的相对定量表达。通过CCK - 8检测法检测细胞增殖,而用Transwell检测法测定细胞的迁移能力。进行毛细血管样管形成检测以检测细胞的管形成能力。在此之后,通过qRT - PCR和蛋白质印迹分析确定miR - 195 - 5p和VEGF - A的相对表达。实验结果表明,与对照组相比,HUVECs和CRNV组织中SNHG1和VEGF - A水平上调,而在CRNV组织和HUVECs中miR - 195 - 5p水平下调,表明lncRNA与miRNA之间存在负相关。过表达的血管内皮生长因子促进细胞增殖和管形成;然而,其沉默导致血管生成和增殖受到抑制。SNHG1的潜在结合位点显示miR - 195 - 5p是其直接靶点,且SNHG1作为该miRNA的海绵。SNHG1的敲低和下调水平显示CRNV细胞的血管生成和迁移显著减少和受到抑制。研究表明,SNHG1抑制显著降低了转染lncRNA SNHG1的HUVECs中的细胞增殖、迁移和管形成。对SNHG1的机制性研究表明,SNHG1作为miR - 195 - 5p的海绵并上调VEGF - A的水平。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/0c29/8548121/7e116beb75f3/joph2021-6646512.001.jpg

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