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. 2020 Mar 4;10(9):3967–3979. doi: 10.7150/thno.40566

Figure 4.

Figure 4

MiR-146a-5p targets the 3′-UTR of TRAF6 and suppresses its expression. (A) TRAF6 was identified as the putative target of miR-146a-5p in five miRNA target prediction algorithms. (B-D) qPCR and Western blot assays were used to detect TRAF6 mRNA and protein levels in PDAC cells transfected with miR-146a-5p mimics, inhibitors or their NCs. (E) A schematic showing the sequence alignment of wild-type (WT) and mutant (MU) miR-146a-5p target sites on the 3′-UTR of TRAF6. Both the WT and MU 3'-UTRs were cloned into luciferase reporter constructs. (F) Dual-luciferase analysis of cotransfection of miR-146a-5p with the WT or MU 3′-UTR of TRAF6 in PDAC cells (n.s, not significant, **P < 0.01). (G) TRAF6 expression in tumor and adjacent normal tissues from 36 PDAC patients was determined by qPCR. (H) The relationship of the expression between miR-146a-5p and TRAF6 in 36 PDAC tissues (P = 0.0017). (I-J) Tissue microarray (TMA) analysis by IHC staining showed that increased TRAF6 protein expression was observed in 87 PDAC tissues (P < 0.0001). (K-L) Kaplan-Meier analysis showed that increased TRAF6 expression was significantly associated with poorer OS and DFS in 87 PDAC patients.