SiRNA silencing of VEGF, IGFs, and their receptors in human retinal microvascular endothelial cells

American Journal of Translational Research

Charles L Cai¹  Fayez Bany-Mohammed²  Yona Nicolau³  Jacob V Ara⁴ 

[1] Department of Ophthalmology, State University of New York, Downstate Medical Center, Brooklyn, NY 11203, USA;Department of Pediatrics, Division of Neonatal-Perinatal Medicine State University of New York, Downstate Medical Center, Brooklyn, NY 11203, USA;Department of Pediatrics, Division of Neonatal-Perinatal Medicine, University of California Irvine, Irvine, CA 92868, USA;State University of New York Eye Institute, New York, NY 10075, USA

关键词: Angiogenesis;    insulin-like growth factor;    retinal endothelial cells;    small interfering RNAs;    vascular endothelial growth factor;   

DOI  :  

学科分类：医学（综合）

来源: e-Century Publishing Corporation

【 摘 要 】

Vascular endothelial growth factor (VEGF) is a potent mitogen that regulates proliferation, migration, and tube formation of endothelial cells (EC). VEGF has recently become a target for severe retinopathy of prematurity (ROP) therapy. We tested the hypothesis that a specific VEGF isoform and/or receptor acts synergistically with insulin-like growth factor (IGF)-I to alter normal retinal microvascular EC angiogenesis and RNA interference can be used to reverse VEGF effects. We used small interfering RNA (SiRNA) transfection to target VEGF isoforms, IGFs, and their receptors in human retinal microvascular endothelial cells (HRECs). Media was collected at 24 and 48 hours post transfection for measurement of VEGF, sVEGFR-1 and IGF-1 levels; and HRECs were assessed for migration, tube formation, VEGF signaling genes, oxidative stress, and immune-reactivity. At 24 hours post transfection VEGF increased with VEGFR-2; sVEGFR-1 decreased with VEGF165, VEGFR-2, and IGF-1R; and IGF-I increased with VEGF189, VEGFR-1, IGF-2R, IGF+VEGF165, and IGF+VEGF121. IGF-I transfection with each VEGF isoform reduced sphere- forming and migration capacities with robust upregulation of caspase-9, COX-2, MAPK, PKC, and VEGF receptors. At 48 hours, the effects were reversed with a majority of genes downregulated, except with IGF-I and NP-1 transfection. Using RNA interference for targeted inhibition of VEGF isoforms in conjunction with IGF-I may be preferable for suppression of HREC overgrowth in vasoproliferative retinopathies such as ROP.

【 授权许可】

CC BY-NC