Figure 3.

Thymidylate synthase (TYMS) is a direct target of microRNA (miR)‐375‐3p in colorectal cancer. A, Alignment of the potential miR‐375 binding site in the 3′‐UTR of TYMS mRNA. B‐E, mRNA (B, C) and protein levels (D, E) of TYMS were measured by quantitative real‐time PCR and western blotting, respectively, in HT29 and HCT116 cells transfected with miR‐375‐3p mimics. F, Protein levels of TYMS were measured by western blotting in HT29 and HCT116 cells transfected with an miR‐375‐3p inhibitor. G, Luciferase assay of HT29 cells transfected with a dual‐luciferase reporter vector containing WT or mutant 3′‐UTR of TYMS mRNA. At 24 h after transfection, cells were further transfected with miR‐375‐3p or miR‐NC (negative control), and the luciferase activity was measured. Cells were treated with 100 nmol/L miRNA and 1 μg/mL 5‐fluorouracil (5‐FU) concentration. All data are shown as the means ± SEM of 3 independent experiments. **P < .01, ***P < .001, ****P < .0001