Figure 5
From: microRNA-139-5p exerts tumor suppressor function by targeting NOTCH1 in colorectal cancer
NOTCH1 is a direct target of miR-139-5p in colon cancer. (A) Human NOTCH1 3’UTR binding site for miR-139-5p. (B) The miR-139-5p wild type binding sequence or its mutated form was inserted into C-terminal of the luciferase gene to generate pMIR-NOTCH1-3’UTR or pMIR-NOTCH1-mut-3’UTR, respectively. (C) miR-139-5p targeted the wild-type but not the mutant 3’UTR of NOTCH1. The data are means ± SD. (D) Ectopic expression of miR-139-5p downregulated NOTCH1 mRNA expression in DLD1 and HCT116 cells as determined by quantitative RT-PCR. (E) miR-139-5p decreased NOTCH1 protein level in DLD1 and HCT116 by western blot. (F) miR-139-5p significantly suppressed NOTCH1 downstream effectors including hairy and enhancer of split-1 (HES1), cyclin D1 and FADD-like apoptosis regulator (CFLAR) transcription. (G) Expression of NOTCH1 is significantly upregulated in primary colorectal tumors compared with their adjacent normal tissues (n = 45). (H) Scatter plots showing the inverse association between miR-139-5p level and NOTCH1 mRNA expression.