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Figure 4 | Molecular Cancer

Figure 4

From: Loss of miR-638 in vitro promotes cell invasion and a mesenchymal-like transition by influencing SOX2 expression in colorectal carcinoma cells

Figure 4

miR-638 suppresses SOX2 expression by directly targeting its 3′-UTR. A) The luciferase activity of the luciferase reporter plasmid containing the 3′-UTRs of four putative miR-638 target genes was assessed using the Dual Luciferase Reporter Gene Assay. After transfection with miR-638 mimics for 48 hours, the relative luciferase activity of the plasmid containing the SOX2 3′UTR was inhibited by 45% by miR-638, and the relative luciferase activity of other plasmids exhibited no significant change. Each sample was compared to the negative control (empty vector), and cotransfection of a Renilla plasmid served as an internal control. B) A potential miR-638 binding site and the mutated sequence in the SOX2 3′UTR for the seed region are shown. C) The luciferase activity of the reporter vector containing either the wild-type (WT) or mutant (MT) SOX2 3′-UTR was assessed after cells were transfected with miR-638 mimics; the mutant abolished the repression by transfection with the miR-638 mimic. (D) Changes in miR-638 expression levels drove endogenous SOX2 mRNA expression changes inversely. (E) miR-638 expression changes drove SOX2 endogenous SOX2 protein expression changes inversely. F) miR-638 expression was inversely correlated with SOX2 mRNA expression in CRC tissues. Based on the miR-638 expression levels in the CRC tissue samples, we sorted these 36 samples into low- and high-expression groups (unpaired t-test).

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