miR-17-5p regulates aniogenesis in vitro. A: Real-time PCR detected miR-17-5p expression after HUVECs transfected with plasmids. B: Cell viability of HUVECs was examined using CCK-8 assay after transfected with miR-17-5p inhibition plasmid, miR-17 plasmid and negative control. C: Ki-67 expression in HUVECs was detected by Immunofluorescence analysis. The bottom line was the amplification of the important parts of Merge. D: Cell cycle analysis showed the viability of HUVECs after altered miR-17-5p expression. E: Migration ability of HUVECs was analyzed using transwell assays. F: Representative images of capillary-like structure formed by HUVECs on Matrigel. The bottom line was the amplification of the capillary-like structure.

miR-17-5p regulates aniogenesis in vivo. (A-B): Morphology of SIVs in 5 dpf Tg(fli1a:EGFP) embryos injected with miR-17 Morpholino and negative control. The bottom line was the amplification of SIVs. The data shown were representative of at least three independent experiments. * p < 0.05. ** p< 0.01.

MiR-17-5p knockdown suppressed the proliferation of CNE2. A: MiR-17-5p expression in 10 NPC tissues and 5 normal nasopharyngeal tissues by Real-time PCR analysis. B: Relative miR-17-5p expression in NPC cell lines and normal nasopharyngeal epithelial cell line. U6 was an endogenous control. (C-D): Immunofluorescence analysis and Real-time PCR analysis of miR-17-5p expression after CNE-2 cells transfected with plasmids. E: Proliferation of CNE-2 cells was examined using CCK-8 assay after transfected with miR-17-5p inhibition plasmid, miR-17 plasmid and negative control. F: Cell cycle analysis by flow cytometry in CNE-2 cells containing different levels of miR-17-5p. The data shown were representative of at least three independent experiments. * p < 0.05. ** p< 0.01.

MiR-17-5p knockdown suppressed the migration of CNE-2. A: Using transwell assay to detect the penetration of CNE-2 cells transfected with plasmids through the membrane comparing with the control. B: Western blot showed the expression of EMT markers after CNE-2 cells transfected with miR-17-5p-specific plasmids and a negative control plasmid. β-actin was the loading control.

HUVECs ingested NPC derived exosomal miR-17-5p to promote angiogenesis. A: Real-time PCR analysed miR-17-5p expression in HUVECs which was co-cultured with CM from CNE-2 cells. B: Representative electron microscopy image of exosomes. C: Western blot showed the level of different kinds proteins in exosomes and CNE-2 cells. Flotillin-1 was the loading control. D: MiR-17-5p expression in exosomes was detected by Real-time PCR. E: Confocal microscopy images showed the ingestion of PKH67-labeled exosomes by HUVECs. Hoechst stained the nuclei (blue) and PKH67 labeled the exosomes (green). F: Real-time PCR showed miR-17-5p expression in HUVECs after absorbed exosomes from CNE-2 cells. (G-H): Morphology of SIVs in 5 dpf Tg(fli1a:EGFP) embryos injected with CNE-2 derived exosomes.

miR-17-5p targeted BAMBI expression and regulated AKT/VEGF-A signaling. A: Wild-type miR-17-5p target sequences of BAMBI mRNA 3'-UTR. Using luciferase reporter assays to quantitatively detect the relative luciferase activities of wild-type and mutant. (B-D): Quantifications of BAMBI mRNA and protein level in HUVECs using Real-time PCR and western blot. E: Real-time PCR detected BAMBI expression in HUVECs incubated with CNE-2 derived exosomes. F: Human VEGF-A Precoated ELISA Kit was used to measure serum VEGF-A levels in 6 NPC patients and 6 healthy controls. G: Western blot of BAMBI, p-AKT, AKT and VEGF-A expression in HUVECs. β-actin as the loading control. The data shown were representative of at least three independent experiments. * p < 0.05. ** p< 0.01.