Figure 5.

ESCC-secreted exosomal miR-320b targets PDCD4 to induce lymphangiogenesis

(A) Schematic representation of the putative predicted miR-320b targeting site in the PDCD4 mRNA 3′ UTR region. (B) Relative reporter gene activity of psiCHECK2 vector bearing PDCD4 3′ UTR fraction in 293T cells co-transfected with increasing amounts (1, 5, and 10 pmol) of miR-320b mimic. (C) Reporter gene activity of psiCHECK2 vector bearing PDCD4 3′ UTR wild-type (WT) fraction or their mutant type (Mut) fraction in 293T cells in the presence of 10 pmol miR-320b mimics. Treatment with PBS, KYSE150-EXO_Vector, or KYSE150-EXO_miR-320b on the levels of PDCD4 RNA (D) and PDCD4 protein (E) in HLECs. (F) Protein levels of PDCD4 and AKT pathway were, respectively, detected by western blot in HLECs treated with indicated exosomes in the presence of PDCD4 overexpression plasmid or vector. Representative images (G) and histogram analysis (H) of tube formation and migration by HLECs treated with KYSE150-EXO_Vector or KYSE150-EXO_miR-320b and then transfected with vector or PDCD4 plasmid. Scale bars, 100 μm. (I) ELISA assay shows the expression level of VEGF-C in the ESCC cells culture medium of different groups. ∗∗p < 0.01, ∗∗∗p < 0.001.