Figure 2. miR-483-5p and miR-551a suppress colorectal cancer cells survival and metastasis in the liver through regulation of CKB.
A, Expression of CKB in SW480 cells whose endogenous miR-483-5p or miR-551a was inhibited with LNAs. B, Liver metastasis in mice injected intrasplenically with 5 × 105 control SW480 cells and CKB over-expressing SW480 cells (n=5). Mice were euthanized at 28 days after injection. C, Liver metastasis in mice injected intrasplenically with 5 × 105 LvM3b expressing a control hairpin or two independent shRNA hairpins targeting CKB (n=6). Mice were euthanized 21 days after injection. D, Survival of control SW480 and CKB over-expressing SW480 cells in organotypic liver slices (n=8). E, Survival of LvM3b cells expressing a control hairpin or hairpin targeting CKB in organotypic slice cultures (n=8). Representative images at day 0 and day 2 are shown. Scale bar represent 50um. F, Relative in vivo caspase activity of control or CKB over-expressing SW480 cells Caspase activity was measured on day 1, 4 and 7 after injection (n=3). G, Relative in vivo caspase-3 activity of SW480 cells expressing a control shRNA or shRNA targeting CKB. Caspase activity was measured on day 1, 4 and 7 after injection (n=3). H, Liver metastasis in mice injected with 5 × 105 SW480 cells whose endogenous miR-483-5p or miR-551a was inhibited by LNA, with and without CKB knockdown. Error bars, s.e.m; all P values are based on one-sided Student’s t-tests, or where appropriate, Mann-Whitney test for non-Gaussian distribution. *p<0.05; **p<0.01; ***p<0.001. See also Figure S2.