Research Paper Volume 13, Issue 17 pp 21435—21450

MiR-185-5p targets RAB35 gene to regulate tumor cell-derived exosomes-mediated proliferation, migration and invasion of non-small cell lung cancer cells

Knockdown of RAB35 reduces the number of exosomes from tumor cells and inhibits its function. The miR-185-5p mimic, RAB35 siRNA and their negative control were transfected into A549 and H2170 cells, respectively. Next, the cell culture medium was collected after 48 hours and extracted exosomes. (A–C) Western blotting was used to detect the relative protein expression of TSG101, CD63 and HSP70 in A549 cells, H2170 cells and their secreted exosomes, and the results suggested that exosomes isolated from A549 and H2170 cells transfected with miR-185-5p mimic and siRAB35 had lower protein expression of TSG101, CD63 and HSP70. β-actin was used as an invariant internal control for calculating protein-fold changes. N=6, ** P

Figure 7. Knockdown of RAB35 reduces the number of exosomes from tumor cells and inhibits its function. The miR-185-5p mimic, RAB35 siRNA and their negative control were transfected into A549 and H2170 cells, respectively. Next, the cell culture medium was collected after 48 hours and extracted exosomes. (AC) Western blotting was used to detect the relative protein expression of TSG101, CD63 and HSP70 in A549 cells, H2170 cells and their secreted exosomes, and the results suggested that exosomes isolated from A549 and H2170 cells transfected with miR-185-5p mimic and siRAB35 had lower protein expression of TSG101, CD63 and HSP70. β-actin was used as an invariant internal control for calculating protein-fold changes. N=6, ** P<0.01.