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We previously demonstrated that vascular endothelial cells released VEGF-enriched exosomes to promote the tumor vasculogenesis and progression after anti-angiogenic therapies (AATs). To clarify how microRNA (miR)-9 promoted the angiogenesis of tumor-associated endothelial cells, in the present study, we investigated the association between miR-9 and sphingosine-1-phosphate (S1P) receptors in angiogenesis. The levels of miR-9 and S1P receptors in normal and tumor endothelial cells were compared with EndoDB database and their correlations were analyzed. The levels of S1P 1 , S1P 2 , and S1P 3  were detected in miR-9 overexpressing endothelial cells by qRT-PCR and western blot. The binding sites of miR-9 on S1P 1  and S1P 3  were predicted and tested by dual-luciferase reporter assays. Then, angiogenesis in endothelial cells overexpressing both S1P 1  and miR-9 was detected. The results showed that miR-9 is overexpressed in ECs from medulloblastoma and glioblastoma xenograft, which is negatively associated with S1P 1  and S1P 3 . Overexpression of miR-9 significantly inhibited S1P 1  and S1P 3  in both mRNA and protein levels. We predicted that binding sites exist between miR-9 and S1P 1 , S1P 3 , but only S1P 1  was directly targeted by miR-9. Overexpression of S1P 1  significantly suppressed the miR-9-induced angiogenesis. Therefore, miR-9 induces angiogenesis via targeting on S1P 1 .

MiR-9 Promotes Angiogenesis via Targeting on Sphingosine-1- Phosphate Receptor 1