Figure 3

Ectopic expression of miR-181a enhanced CRC cell in vitro migration and invasion as well as in vivo liver metastasis. (A) The expression of miR-181a/b normalized to U6 was assayed in pri-miR-181a lentivirus infected HT-29 cells (HT29-181a), HT29 cells infected with control vector (HT29-NC) and parental HT29 cells (HT29). The data represent fold changes in miRNA expression. The limits of the 95% confidence intervals indicate variability of the changes; (B) Cell growth rates were stimulated by miR-181a in HT29 cells detected by CCK-8 assay. (C) The effect of miR-181a on the growth of HT-29 cells was evaluated in NOD-SCID mice. (C-1) Tumour growth curves. Points and bars represent the mean tumor volume with SD, eight mice in each group, p < 0.05; (C-2) Tumor weight derived from HT-29 cells expression different levels of miR-181a; (C-3) Tumors derived from HT-29 cells expression different levels of miR-181a; (D) Wound-healing assay shows that cell motility was enhanced following miR-181a over-expression in HT29 cells. D-1) Representative images from wounds made using a confluent monolayer of HT-29, HT29-NC and HT29-181a cells taken at 0 h and 48 h post wounding, imaged using IncuCyte ZOOM. D-2) The quantification results of cell motility (wound confluence) are plotted. Data represent the mean ± SD of three independent experiments. (E) Boyden chamber assays were performed with and without matrigel. The quantification results of migrated cells and invaded cells through Matrigel are plotted in (E-2) and (E-3), respectively. Data represent the mean ± SD of three independent experiments with six random fields counted for each chamber; (F) Wound-healing assay shows that cell motility was enhanced following miR-181a over-expression in SW480 cells. (G) Cell migration and invasion were significantly enhanced in SW480 overexpressing miR-181a cells. (H) miR-181a enhances HT-29 cells liver metastasis. Representative livers derived from NOD-SCID mice are shown.