MicroRNA-323a-3p Negatively Regulates NEK6 in Colon Adenocarcinoma Cells

Figure 2

miR-323a-3p negatively regulates NEK6. (a) The miRNAs targeting NEK6 predicted from TargetScan, MicroT-CDS, MiRDB, StarBase, and TarBase were drawn in Venn diagrams. (b) The 3′ UTR of the NEK6 and miR-323a-3p binding sites and the 3′ UTR mutation sites (red) of NEK6 are shown. (c) miR-323a-3p was demonstrated to target the 3′ UTR of NEK6 using a dual-luciferase reporter assay. (d–f) After miR-323a-3p was overexpressed in HCT-8 and SW620 cells, RT-PCR (d, e) and western blotting (f) assays were employed to assess the expression of miR-323a-3p and NEK6. (g) The miR-323a-3p level in COAD tissues from public databases was analyzed. (h) miR-323a-3p levels in COAD cell lines were detected via RT-PCR. (i) The correlation between miR-323a-3p and the overall survival of COAD patients was evaluated via the KM curve. 3′ UTR WT: the wild-type 3′ noncoding region of NEK6; 3′ UTR MUT: the mutant 3′ noncoding region of NEK6; ns: not significant. and .

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