Fig. 4

Interaction between N-BLR and miR-200 family members. a The effect of transient transfection of N-BLR siRNA3 and siRNA4 on the miR-200 family in Colo320 cells. MiR-141-3p and miR-200c-3p were increased in both N-BLR siRNAs transfected cells compared with scramble control. b A luciferase vector including the full N-BLR sequence (pGL3-N-BLR) as well as vectors that were mutated separately at the interaction sites of either miR-141-3p or miR-200c-3p [pGL3-N-BLR(M)] were constructed. Luciferase activity is decreased only when miR-141-3p and miR-200c-3p are co-transfected with the WT construct but not when a mutated vector is used. c Most representative images from ISH of tissue microarray showed lower levels of both miR-141-3p and miR-200c-3p in adenocarcinoma tissue compared with normal tissue, whereas an inverse pattern was found for N-BLR levels. d Image analysis were performed to evaluate the association between the levels of miR-141-3p and miR-200c-3p and those of N-BLR. The quantification was performed in a pair-matched fashion, so that the levels of the three targets were quantified on the same tissue spot of the microarray. e N-BLR and E-cadherin expression in tumor and normal samples: N-BLR was increased and E-cadherin was decreased in CRC when compared to normal colon. f The same is true when CRC with lymph node invasion (LN+) were compared with cases without lymph node involvement (LN–). Asterisks mark cases with statistically significant difference compared with scrambled. (Student’s t-test; *p < 0.05; **p < 0.01; ***p < 0.001; ****p < 0.0001)