MicroRNA-210 Modulates Endothelial Cell Response to Hypoxia and Inhibits the Receptor Tyrosine Kinase Ligand Ephrin-A3

Abstract

MicroRNAs (miRNAs) are small non-protein-coding RNAs that function as negative gene expression regulators. In the present study, we investigated miRNAs role in endothelial cell response to hypoxia. We found that the expression of miR-210 progressively increased upon exposure to hypoxia. miR-210 overexpression in normoxic endothelial cells stimulated the formation of capillary-like structures on Matrigel and vascular endothelial growth factor-driven cell migration. Conversely, miR-210 blockade via anti-miRNA transfection inhibited the formation of capillary-like structures stimulated by hypoxia and decreased cell migration in response to vascular endothelial growth factor. miR-210 overexpression did not affect endothelial cell growth in both normoxia and hypoxia. However, antimiR-210 transfection inhibited cell growth and induced apoptosis, in both normoxia and hypoxia. We determined that one relevant target of miR-210 in hypoxia was Ephrin-A3 since miR-210 was necessary and sufficient to down-modulate its expression. Moreover, luciferase reporter assays showed that Ephrin-A3 was a direct target of miR-210. Ephrin-A3 modulation by miR-210 had significant functional consequences; indeed, the expression of an Ephrin-A3 allele that is not targeted by miR-210 prevented miR-210-mediated stimulation of both tubulogenesis and chemotaxis. We conclude that miR-210 up-regulation is a crucial element of endothelial cell response to hypoxia, affecting cell survival, migration, and differentiation.

The regulatory role of microRNAs in angiogenesis‐related diseases

“…MiR‐210 promotes angiogenesis and the maturation of vasculature in post‐ischaemic brain tissue by enhancing the expression of Notch, VEGF and vascular endothelial growth factor receptor‐2 (VEGFR‐2) in HUVECs 90. Further research indicated that miR‐210 facilitates angiogenesis through negatively regulating the target gene, ephrin A3, which is an important member of the ephrin angiogenesis regulatory gene family 51, 91, 92. Moreover, a study revealed that isoprenaline enhances the levels of miR‐210, miR‐21 and miR‐1 and decreases those of the lncRNAs maternally expressed 3 (MEG3) and growth arrest‑specific transcript 5 (GAS5), thereby improving angiogenesis 93.…”

Extracellular vesicles swarm the cancer microenvironment: from tumor–stroma communication to drug intervention

“…For example in colorectal cancer, tumour-derived EVs can promote proliferation of endothelial cells and enhance their cell-cycle activities through M-phase related mRNAs, such as those coding for the centromere protein E (CENPE), PDZ binding kinase (PBK) or cyclin-dependent kinase 8 (CDK8) 32 . Additionally, the involvement of vesicular miRNAs in neo-angiogenesis has been studied such as miRNA-210 that exhibited strong pro-angiogenic activity 22,40,83 . Furthermore, miRNA-210 has been observed to suppress the expression of specific genes such as EFNA3 (coding for Ephrin-A3) in endothelial cells, resulting in enhanced neo-angiogenesis 21,39,73 .…”

hsa‐miR‐210 is a marker of tumor hypoxia and a prognostic factor in head and neck cancer

“…13 HIF-1alpha induces microRNA-210 (hsa-miR-210) directly through its interaction with its promoter sequence. Validated targets of hsa-miR-210 include the receptor tyrosine kinase ligand ephrin-A3, 14 the transcription factor E2F3, 15 and the DNA repair enzyme RAD52. 16 However, the effect of hsa-miR-210 may depend on the cancer type; whereas high level of expression is an independent prognostic factor in breast cancer, 17 it is frequently deleted in epithelial ovarian cancer.…”