Fig. 5

Exosome-delivered MiR-126 from endothelial cells as a treatment of malignant nasal-septum carcinoma (MNSC) cells. a Cancer cell-stromal cell cross talk. Triple co-culture was performed by layering fibroblasts (IMR-90) and endothelial cells (HUVEC) on two opposite surfaces of trans-well inserts, and MNSC cells were cultured at the bottom of the trans-well. The co-culture was treated with exosomes from HUVEC (exo-HUVEC, 50 μg/ml), and after 2 days of treatment MiR-126 level and the expression of its gene targets were evaluated in MNSC cells. b Effect of exosomes from endothelial cells on colony-forming activity in MNSC cells. The images shown represent 3 months of treatment (magnification 2.5×). c Immunoblot evaluation of MAPK signalling. MNSC cells in triple co-culture were treated with exo-HUVEC (20 μg/ml), and after 2 days of incubation evaluated for the expression of IRS1, pERK1/2, ERK1/2, p-p38, p38 (upper-panel). Densitometry evaluation of the bands related to the level of actin (down-panel). d Representative images of wound healing mobility assay (left-panel) and proliferative index Ki67 staining (right-panel). MNSC cells in triple co-culture were treated with exo-HUVEC (20 μg/ml), and after incubation evaluated for wound-healing mobility percentage and percentage of Ki67 positive cells. All experiments were carried out in triplicate. The symbol “*” denotes significant differences between non treated cells (CTRL) and exo-HUVEC treated cells, p < 0.05