MiR-126 overexpression inhibits high glucose-induced migration and tube formation of rhesus macaque choroid-retinal endothelial cells by obstructing VEGFA and PIK3R2

• Published in: Journal of diabetes and its complications Vol. 31; no. 4; pp. 653 - 663
• Main Authors: Yang, Wenzhi, Yang, Jin, Xue, Liping, Xiao, Libo, Li, Yan
• Format: Journal Article
• Language: English
• Published: United States Elsevier Inc 01-04-2017; Elsevier Limited
• Subjects: Angiogenesis; Animals; Breast cancer; Cancer therapies; Cell Line; Cell Movement; Choroid - metabolism; Choroid - pathology; Class Ia Phosphatidylinositol 3-Kinase - antagonists & inhibitors; Class Ia Phosphatidylinositol 3-Kinase - chemistry; Class Ia Phosphatidylinositol 3-Kinase - genetics; Class Ia Phosphatidylinositol 3-Kinase - metabolism; Computational Biology; Diabetes; Diabetes mellitus; Diabetic retinopathy; Diabetic Retinopathy - metabolism; Diabetic Retinopathy - pathology; Endocrinology & Metabolism; Eye Proteins - agonists; Eye Proteins - antagonists & inhibitors; Eye Proteins - genetics; Eye Proteins - metabolism; Gene Expression Regulation; Gene Knockdown Techniques; Genes, Reporter; Glucose; Glucose - metabolism; Hyperglycemia - metabolism; Hyperglycemia - pathology; Lasers; Macaca mulatta; MicroRNAs; MicroRNAs - antagonists & inhibitors; MicroRNAs - chemistry; MicroRNAs - metabolism; miR-126; Molecular Mimicry; Osmolar Concentration; Pathogenesis; PIK3R2; Retina - metabolism; Retina - pathology; RNA - metabolism; Roles; Signal Transduction; Stereoisomerism; Vascular endothelial growth factor; Vascular Endothelial Growth Factor A - agonists; Vascular Endothelial Growth Factor A - antagonists & inhibitors; Vascular Endothelial Growth Factor A - genetics; Vascular Endothelial Growth Factor A - metabolism; VEGFA; miR-126; diabetes mellitus; PIK3R2; diabetic retinopathy; VEGFA; Diabetic retinopathy; Diabetes mellitus
• ISSN: 1056-8727; 1873-460X
• DOI: 10.1016/j.jdiacomp.2016.12.004

Abstract Aim The aims of this study are to investigate the relative regulation between miR-126 and VEGF/PI3K/AKT signaling pathway in retinal vascular endothelial cells. Methods Rhesus macaque choroid-retinal endothelial cell line (RF/6A) cells were cultured in high glucose to imitate the conditions occurring in DR. First, we detected the expression of miR-126, VEGFA and PIK3R2 in RF/6A cells on the condition of high glucose by q-PCR and western blot. Then, after addition of miR-126 mimics and miR-126 inhibitor, we investigated the function of miR-126 in RF/6A cells by scratch wound, transwell migration and tube formation assays, and the effect of miR-126 on the expression of VEGFA, PIK3R2 and AKT. Moreover, bioinformatics analysis and luciferase array were used to confirm the direct or specific regulation of miR-126 to VEGFA or PIK3R2. Results Here, first, we found that high glucose could induce the decrease of miR-126, the increase of VEGFA and PIK3R2 in RF/6A. Then, by scratch wound, transwell migration and tube formation assays, we found that miR-126 overexpression could inhibit the migration and sprouting of RF/6A cells induced by high glucose. While knockdown of miR-126 led to the opposite results. Moreover, overexpression miR-126 inhibited the increased expression of VEGFA, PIK3R2, SDF-1α, VCAM-1, SPRED1, and the activation of AKT1 induced by high glucose and miR-126 inhibitor caused the opposite results which were determined by q-PCR and western blot. In addition, by luciferase assay, we found that miR-126 could directly negative regulate VEGFA and PIK3R2. Conclusion Our results suggest that miR-126 overexpression inhibits the migration and sprouting of RF/6A cells induced by high glucose which might possibly by blocking VEGFA and PIK3R2 in the VEGF/PI3K/AKT signaling pathway.