Fig. 5

Exosomes transfer miR-146a-5p from CAFs to PCa cells. a CAFs transiently transfected with Cy3-tagged miR-146a-5p or with the miR-control, and GW4869-treated CAFs transfected with Cy3-tagged miR-146a-5p were co-cultured with DU145 for 24 h. Fluorescence microscopy was performed to observe the green signals (Vimentin) and red fluorescent signals (Cy3) in DU145 (scale bars = 6.25 μm). b Exosomes were isolated from the supernatants of CAFs transfected with cy3-tagged miR-146a-5p (CAFs-miR-146a-5p-cy3 exo) or with miR-control (CAFs-miR-control exo) and exosomes (25 μg/mL) were added to DU145 for 24 h. Fluorescence microscopy was performed to detect the green signals (Vimentin) and red fluorescent signals (Cy3) in DU145 (scale bars = 6.25 μm). c The expression of miR-146a-5p in LNcaP and DU145 cells were measured using Real-time PCR at 24 h after treating with exosomes (25 μg/mL) derived from DHT/ETOH-treated CAFs. d The expression of miR-146a-5p in lung metastasis of the nude mice among three groups by Real-time PCR. e The level of miR-146a-5p in androgen deprived prostate tissues (Castration) was compared with that of the control PCa tissues (NC) (n = 8) by Real-time PCR. f and g LNcaP cells were co-cultured with CAFs-derived exosomes and transfected with miR-NC or miR-146a-5p mimics for 48 h, and the migration and invasion of LNcaP cells was assessed by a Transwell assay (scale bars = 25 μm). h and i DU145 cells were co-cultured with CAFs-derived exosomes and transfected with miR-NC or miR-146a-5p mimics for 48 h, and the migration and invasion of DU145 cells was assessed by a Transwell assay (scale bars = 25 μm). j and k LNcaP and DU145 cells were co-cultured with CAFs-derived exosomes and transfected with miR-NC or miR-146a-5p mimics for 48 h, and western blotting analysed the protein levels of EMT markers and MMPs. Data are shown as mean ± SD representing triplicate measurements. (Student’s t-test, * P < .05, **P < .01, ***P < .001, ****P < .0001)