Figure 5.
Exosome-transmitted miRNA-335-5p promotes CRC cell invasion, metastasis, and EMT, as well as decreases the level of RASA1
(A) Determination of the migration and invasion abilities of SW480 cells treated with NC-exo (SW480-exo transfected with NC miRNA) or miR-335-5p mimics-exo (SW480-exo transfected with miR-335-5p mimics) using a Transwell assay. Data are presented as mean ± SD (n = 3). ∗∗p < 0.01, ∗∗∗p < 0.001. Scale bars, 100 μm. (B) Determination of the migration and invasion abilities of SW620 cells treated with NC-exo or miR-335-5p mimics-exo using a Transwell assay. Data are presented as mean ± SD (n = 3). ∗∗p < 0.01. Scale bars, 100 μm. (C) Western blot analysis of RASA1 and Ras protein expression levels in SW480 and SW620 cells treated with NC-exo or miR-335-5p mimics-exo. (D) Western blot analysis of E-cadherin and vimentin protein expression levels in SW480 and SW620 cells treated with NC-exo or miR-335-5p mimics-exo. (E) Bioluminescence assay evaluating tumor metastases in mice. SW480 cells transduced with a lentiviral vector encoding the firefly luciferase gene (3 × 106 cells per mouse, n = 6) were injected into the tail vein of NOD/SCID mice, and the in vivo bioluminescent signal was quantified at the 9th week. Representative in vivo images of NC-exo or miR-335-5p mimics-exo treated mice are shown. Data are presented as mean ± SD (n = 6). ∗∗p < 0.01. (F) Representative photographs of macroscopic lung appearance. The number of metastatic tumor nodules for each group is shown on the right panel. Data are presented as mean ± SD (n = 6). ∗∗p < 0.01.