LncRNA VIM-AS1通过miR-497-5p/FBXW7轴调控高糖环境下视网膜内皮细胞的迁移和凋亡

LncRNA VIM-AS1 regulates cell migration and apoptosis of retinal endothelialcells under high glucose treatment via the miR-497-5p/FBXW7 axis

目的:探讨长链非编码RNA(lncRNA)VIM反义RNA 1(VIM-AS1)在糖尿病视网膜病变中的潜在分子机制。方法:使用q RT-PCR测定LncRNA VIM-AS1、miR-497-5p和FBXW7 mRNA的表达。使用蛋白质印迹检测FBXW7蛋白水平。分别使用CCK-8实验、伤口愈合实验和流式细胞技术分析评估细胞增殖、迁移和凋亡。通过双荧光素酶报告基因分析验证lncRNA VIM-AS1、miR-497-5p和FBXW7之间的结合关系。结果:在高糖处理的ARPE-19细胞中,LncRNAVIM-AS1和FBXW7的表达显著降低,而miR-497-5p的表达上调。LncRNA VIM-AS1可以通过竞争性结合miR-497-5p上调FBXW7的表达。LncRNA VIMAS1过表达能够促进HG处理的ARPE-19细胞的增殖和迁移,并抑制细胞凋亡,而miR-497-5p过表达消除了lncRNA VIM-AS1过表达对HG处理的ARPE-19细胞的影响。此外,FBXW7敲低消除了miR-497-5p对HG处理的ARPE-19细胞表型的抑制。结论:lncRNA VIM-AS1可通过调控miR-497-5p/FBXW7轴促进HG处理的ARPE-19细胞增殖和迁移,同时抑制细胞凋亡,提示lncRNA VIM-AS1作为治疗靶点潜力巨大。

Objective:Our study aimed to probe the potential molecular mechanism of long non-coding RNA(lncRNA)VIM Antisense RNA 1(VIM-AS1)in diabetic retinopathy.Methods:LncRNA VIM-AS1,miR-497-5p and FBXW7 mRNA expressions were determined using qRT-PCR.The FBXW7 protein level was also detected using western blotting.The cell viability,migration and apoptosis were evaluated using CCK-8 assay,wound healing assay and flow cytometry analysis,respectively.Additionally,the binding relationships among lncRNA VIM-AS1,miR-497-5p and FBXW7 were verified by dual luciferase reporter assaies.Results:LncRNA VIM-AS1 and FBXW7 expressions were remarkably reduced in HG-treated ARPE-19 cells,while miR-497-5p was upregulated.LncRNA VIM-AS1 could upregulate the expression of FBXW7 by competitively binding to miR-497-5p.LncRNA VIM-AS1overexpression promoted cell proliferation and migration,and inhibited cell apoptosis in HG-induced ARPE-19 cells,while miR-497-5p overexpression abolished the effects of lncRNA VIM-AS1 overexpression on HG-induced ARPE-19 cells.Furthermore,FBXW7knockdown abrogated the effects of miR-497-5p inhibition on cell phenotypes of HG-treated ARPE-19 cells.Conclusion:LncRNA VIM-AS1 could promote the proliferation and migration,while inhibited cell apoptosis of HG-treated ARPE-19 cells by regulation of miR-497-5p/FBXW7 axis,suggesting that lncRNA VIM-AS1 might have great potential as therapeutic target for diabetic retinopathy.