Down-regulation of miR-622 in gastric cancer promotes cellular invasion and tumor metastasis by targeting ING1 gene

Figure 4 MiR-622 binds to the 3’UTR of Inhibitor of growth family, member 1 to post-transcriptionally repress the protein levels. A: Representative nucleotide sequence matches between possible target genes and miRNAs. Putative binding site of miR-622 in the ING1 3’UTR region (detected by TargetScan). Only matched nucleotides with miRNA seed sequences are indicated with vertical lines; B: Effects of miR-622 on ING1 protein levels. HEK 293T cells were transfected with 100 nmol/L of miR-622 precursor or miR-622 inhibitor. Whole-cell lysates were prepared 72 h after transfection and subjected to immunoblot analysis using antibodies against ING1 and glyceraldehyde-3-phosphate dehydrogenase (GAPDH). Relative gray scale values were compared with GAPDH; C: Luciferase reporter assay. A total of 1 × 10⁵ HEK 293T cells were transfected with 100 ng wild-type-UTR-reporter or mutant-UTR-reporter constructs together with 100 nmol/L of miR-622 precursor or miR-622 inhibitor as indicated. ^bP < 0.01. Each bar represents mean values ± SE from three independent experiments. ING1: Inhibitor of growth family, member 1; AS-miR-622: An ambion miRNA inhibitor for miR-622.