Research Paper Volume 11, Issue 19 pp 8068—8084

A circular RNA from APC inhibits the proliferation of diffuse large B-cell lymphoma by inactivating Wnt/β-catenin signaling via interacting with TET1 and miR-888

Figure 4. Circ-APC can sponge miR-888 in DLBCL. (A) The CircInteractome and miRanda online tools identified 12 miRNAs with putative binding sites for both circ-APC and APC. (B) RNA pull-down assay with a control or circ-APC probe in U2932 and TMD8 cells, followed by qRT-PCR analysis of the 12 miRNAs identified above. (C) Schematic diagram of the circ-APC luciferase reporter vector with a wild-type or mutant miR-888 binding site. (D) Luciferase reporter assay in U2932 and TMD8 cells co-transfected with the wild-type or mutant circ-APC luciferase vector and control miRNA or miR-888 mimics. (E) RNA pull-down assay in U2932 and TMD8 cells transfected with wild-type or mutant miR-888 mimics, followed by qRT-PCR analysis of circ-APC expression. (F) qRT-PCR analysis of miR-888 expression after circ-APC overexpression. (G) Cell proliferation rate in stably circ-APC-overexpressing U2932 and TMD8 cells transfected with wild-type or mutant miR-888 mimics. (H) Luciferase reporter assay in control or circ-APC-overexpressing U2932 and TMD8 cells co-transfected with the wild-type or mutant APC 3′-UTR luciferase vector and control miRNA or miR-888 mimics. (I) qRT-PCR analysis of APC expression in control or circ-APC-overexpressing U2932 and TMD8 cells transfected with control miRNA or miR-888 mimics. *p < 0.05, **p < 0.01, ***p < 0.001.