miR-132 promotes retinal neovascularization under anoxia and reoxygenation conditions through up-regulating Egr1, ERK2, MMP2, VEGFA and VEGFC expression.

Retinal neovascularization (RNV) is a prominent pathological angiogenesis, which causes detrimental outcomes in visual functions. Previous literature represents that miR-132 induces angiogenesis in tumor development and ischemic diseases. Considering the important role in angiogenesis, we hypothesized that miR-132 might be involved in RNV. In this study, human retinal microvascular endothelial cells were maintained in hypoxia for indicated time, followed by further incubation in normoxic conditions to establish hypoxia/reoxygenation (H/R) models . mRNA microarray analysis was undertaken to detect alterations in gene profiles in the cells. qRT-PCR and Western blotting were performed to evaluate expression of genes that are closely associated to neovascularization. Results showed that miR-132 expression was increased under hypoxic conditions. Reoxygenation for a limited time (6 h) failed to restore miR-132 expression to basal level. Interference of miR-132 expression via its inhibitor suppressed the cell proliferation under H/R conditions, increasing the apoptosis rate. mRNA microarray analysis revealed that miR-132 is involved in the regulation of vasculature development, blood vessel morphogenesis, and proliferation and migration of microvascular endothelial cells through regulating genes such as early growth response gene 1 (Egr1), extracellular signal-regulated kinase (ERK), metal matrix proteinase (MMP2), vascular endothelial growth factor (VEGF)-A and VEGF-C. qRT-PCR and Western blotting further demonstrated that miR-132 up-regulated their gene and protein expression under H/R conditions. In summary, miR-132 was involved in the development of RNV under H/R conditions, at least partly, through up-regulating Egr1, ERK2, MMP2, VEGFA and VEGFC expression. This finding facilitates the understanding of pathogenic mechanisms of RNV.