MiR-30b suppresses tumor migration and invasion by targeting EIF5A2 in gastric cancer

Figure 1 Expression levels of miR-30b in gastric tissue samples and gastric cell lines. A: MiR-30b expression was determined in 23 pairs of gastric cancer tissues compared with corresponding normal tissues by quantitative RT-PCR. Each sample was analyzed in triplicate and normalized to U6. T: tumor tissues; N: adjacent normal tissues; B: Lower miR-30b expression was observed in gastric cancer cell lines compared to that in GES-1; C: AGS and MGC803 cell proliferation was determined by the CCK-8 assay. Upregulation of miR-30b by transfection with mimic suppressed cell proliferation. Downregulation of miR-30b by transfection with inhibitor promoted cell proliferation. Data are displayed as mean ± SD.

Figure 2 Effect of miR-30b on cell apoptosis. The histograms depict apoptosis of AGS cells and MGC803 cells transiently transfected with miR-30 mimic or control.

Figure 3 Effect of miR-30b on the migration and invasion of AGS and MGC803 cells in a transwell assay. A: Overexpression of miR-30b notably inhibited the migration and invasion of AGS and MGC803 cells; B: The migration and invasion abilities of AGS and MGC803 cells were dramatically increased after miR-30b inhibitor treatment. The bar shows the average ± SD of three independent experiments.

Figure 4 miR-30b decreases eukaryotic translation initiation factor 5A2 expression by targeting its 3’-UTR. A: The binding site of miR-30b in the 3’-UTR of eukaryotic translation initiation factor 5A2 (EIF5A2) is highly conserved across species; B: The putative binding sites for miR-30b was found in the 3’-UTR of EIF5A2 at 3664-3671bp; C: miR-30b mimic downregulated luciferase activities controlled by wild-type 3’-UTR of EIF5A2, but did not affect luciferase activity controlled by mutant 3’-UTR of EIF5A2; D: The expression levels of EIF5A2 protein in different gastric cell lines; E: The expression levels of EIF5A2 and of its downstream genes were detected by Western blot analysis in AGS and MGC803 cells transfected with the miR-30b mimic or control for 48 h. β-actin was used as an internal control; F: Western blot analysis of EIF5A2 and its downstream genes transfection of miR-30b inhibitor in AGS and MGC803 cells.