Figure 5

TrkB is a functionally target of miR-204-5p involved in the clonogenic growth, migration and invasion of endometrial cancer cells. (A) Upper panel, Ishikawa cells were co-transfected with miR-204i or its scrambled control and siTrkB or its scrambled control. Expression of TrkB mRNA in Ishikawa cells was measured by RT-PCR assays at 48 h posttransfection. Lower panel, TrkB protein levels were measured by western blotting assays at 72 h posttransfection. Cologenic assays were done as described in Methods and are shown in (B). (C) Migrated and invasive cells on the lower surface of the Transwell filter were stained and photographed, 200×. The number of migrated and invasive cells is shown in (D). (E) Ishikawa cells were transfected with miR-204 m, and the growth curves of Ishikawa cells were plotted by MTT assays. (F) Colony formation was observed. Transwell and cell invasion assays were done as described in Methods. The number of migrated and invasive cells is shown in (G and H). In all the panels, data are shown as mean ± SD of at least three independent experiments. * P < 0.05, ** P < 0.01; NS P > 0.05.