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Fig. 7 | Journal of Experimental & Clinical Cancer Research

Fig. 7

From: Exosomal circTUBGCP4 promotes vascular endothelial cell tipping and colorectal cancer metastasis by activating Akt signaling pathway

Fig. 7

The role of miR-146b-3p in HUVECs, and exosomal circTUBGCP4 contribution to HUVECs dysfunction regulated by miR-146b-3p. (a and b) miR-146b-3p expression in HUVECs treated with HCT116-Exo and SW480-Exo at 0 h, 6 h, 12 h, and 24 h. HUVECs were transfected with miR-146b-3p mimic and NC mimic for 48 h. Then, the migration and tube formation ability of HUVECs were evaluated by transwell migration assay (c), wound healing assay (d), and tube formation assay (e), then wound healing percentage (%) and the number of migrated cells and nodes were analyzed by Image J and Prism 9. (f) Western blot analysis of PDK2, p-AKT (Ser473), AKT and CD34 in HUVECs transfected with miR-146b-3p mimic and NC mimic. GAPDH was the internal control of whole-cell lysates. HUVECs were treated with exosomes derived from LVcircTUBGCP4-Exo (HCT116 and SW480) and its negative control, follow by transfected with miR-146b-3p mimic and NC mimic. After 48 h, the migration and tube formation were measured by transwell migration assay (g) and tube formation assay (h). The number of migrated cells and nodes was analyzed by Image J and Prism 9. (i) Confocal microscopy image of F-actin (red) in HUVECs treated with exosome derived from LVcircTUBGCP4-Exo (HCT116 and SW480) and its negative control, follow by transfected with miR-146b-3p mimic and NC mimic, Scale bars = 10 μm. The yellow dot is used to mark the filopodia represented by F-actin. (j) CD34 immunofluorescence in the above groups, the intensity of CD34 was analyzed by Zeiss Zen software (left), Scale bars = 10 μm. Mean ± SEM. Student's t-test, *P < 0.05, 0.001 < ** P < 0.01, *** P < 0.001

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