The role of lncRNA NEAT1-miR-542-3p-ANG2 regulatory network on Choroidal Neovascularization in Tree Shrew.

Abstract

Choroidal neovascularization (CNV) is a common fundus lesion that can disrupt the normal structure and function of the retina, thereby affecting vision. If left untreated, it can cause irreversible visual loss. The study aimed to investigate the regulatory role of the long non-coding RNA NEAT1-microRNA-542-3p-Angiopoietin-2 (lncRNA NEAT1-miR-542-3p-ANG2) in the development of CNV. Following establishment of a laser-induced CNV model, we knocked down/overexpressed lncRNA NEAT1 and miR-542-3p by intravitreal injection of adenovirus and assessed the outcomes of interference and overexpression by real time quantitative polymerase chain reaction (RT-qPCR). We observed vascular lesions of the retina by hematoxylin-eosin (HE) stain and choroidal flatmounts and detected the expression of angiogenesis-related factors by western blot. Human vascular endothelial cells (HUVECs) were utilised to explore the in vitro effects of lncRNA NEAT1-miR-542-3p-ANG2. The tube formation assay, cell counting kit-8 (CCK-8) assay and wound healing assay were employed to evaluate the angiogenesis, proliferation and migration ability of HUVECs. The expression level of related factors were detected using RT-qPCR and western blotting. Moreover, the interaction between lncRNA NEAT1, miR-542-3p, and ANG2 was assessed using a dual-luciferase reporter assay and RNA pull down assay. NEAT1 knockdown or miR-542-3p overexpression prevent the CNV development and reduced the angiogenesis-related factors levels. And NEAT1 knockdown reduced the angiogenesis, proliferation and migration of HUVECs, as well as angiogenesis-related factor levels, however, these effects were reversed by the presence of the miR-542-3p inhibitor. And miR-542-3p regulated HUVECs function by targeting ANG2. NEAT1 functioned as a competing endogenous RNA (ceRNA) to regulate the expression and function of ANG2 through competitively binding with miR-542-3p, then further affected the formation and development of CNV.