miR-1271-5p negatively regulates CD164 expression by binding to 3′-UTR of CD164. (A) Schematic representation of the potential miR-1271-5p target sequences in CD164 mRNA 3′-UTR predicted by TargetScan. (B) HCT-116 cells were transfected with luciferase reporter plasmid containing wild-type CD164-3′-UTR (CD164-Wt) or mutant CD164-3′-UTR (CD164-Mut), together with miR-1271-5p mimics or negative control miR-NC. The relative luciferase activity was measured 48 h after transfection. (C) The expression of CD164 in the CC tissues and non-tumor control tissues was examined by immunohistochemical staining. Scale bar: 200 µm. (D) The relative expression of CD164 in CC tissues or normal control tissues was evaluated by qPCR analysis. (E) The relative expression of CD164 in CC cell lines (HCT-116, SW620 and DLD-1) or the control cell line FHC was evaluated by RT-qPCR analysis. (F) The relative expression of CD164 in HCT-116 or SW620 cells transfected with miR-1271-5p mimics or miR-NC was evaluated by RT-qPCR analysis. (G) HCT-116 or SW620 cells were transfected with control, sh-RP11-619L19.2, miR-1271-5p inhibitor or sh-RP11-619L19.2 + miR-1271-5p inhibitor. The relative expression of CD164 was evaluated by RT-qPCR analysis 48 h later. *P<0.05, **P<0.01, ***P<0.001. UTR, untranslated region; RT-qPCR, reverse transcription-quantitative PCR.