Fig. 3: MAPK1 and Snail are direct targets of miR-22. | Cell Death & Disease

Fig. 3: MAPK1 and Snail are direct targets of miR-22.

From: MiR-22 suppresses epithelial–mesenchymal transition in bladder cancer by inhibiting Snail and MAPK1/Slug/vimentin feedback loop

Fig. 3

a Venn diagrams of calculating the intersections of the four target prediction engines (miRanda, starBase v2.0, TargetScan and PITA; analyzed by R package VennDiagram). Potential miR-22 target genes were determined as being predicted by at least three of the four target prediction engines (n = 540). b Gene ontology enrichment analysis indicated that these 540 potential targets were involved in cancer and cell motility-related pathways. c qRT-PCR analysis. UM-UC-3 cells were treated with 50 nM miR-22 mimics or NC for 48 h. A decrease in the expression of MAPK1, Snail, TGFBR1, PDGFC and CDKN1A was detected in miR-22 transfected UM-UC-3 cells. d Dual-luciferase reporter assay. MiR-22 effectively suppressed the luciferase activity of vectors that carried 3′-UTRs of MAPK1 and Snail but not TGFBR1, PDGFC, and CDKN1A. e Representative images of the western blotting assay. MiR-22 (50 nM, 100 nM) significantly inhibited the expression of p-ERK2 (p-MAPK1), ERK2 (MAPK1) and Snail in T24 and UM-UC-3 cells. f Schematic representation of the miR-22 predicted binding sites in the 3′-UTRs of MAPK1/Snail mRNAs and 3′-UTR-mutated alignments. g Dual-luciferase reporter assay. The luciferase activities of the mutated vectors of MAPK1 and Snail were unaffected by the transfection of miR-22. Error bars represent the S.D. obtained from three independent experiments. *P < 0.05

Back to article page