Journal: Oncology Reports
Article Title: MicroRNA-92a promotes cell proliferation, migration and survival by directly targeting the tumor suppressor gene NF2 in colorectal and lung cancer cells
Figure Lengend Snippet: NF2 is downregulated by miR-92a-3p via its 3′UTR. (A) Schematic representation of the full and cloned 3′UTR region of NF2 isoform I mRNA and the predicted miR-92a-3p MREs within its sequence. Dual-Luciferase assay of (B) HCT116 and (C) A549 cells transfected with empty vector (i.e., pmirGLO) or NF2 −3′UTR-wild-type. (D and E) The expression levels of mature miR-92a-3p were measured by QuantiGene miRNA assay in untransfected (UTC) or miR-92a construct-transfected (OE) HCT116 and A549 cells. Dual-Luciferase assay of (F) HCT116 and (G) A549 cells co-transfected with increasing plasmid ratios of NF2 −3′UTR-wild-type:miR-92a. (H) Secondary structure analysis of the RNA-RNA interaction (boxed region) between miR-92a-3p (red) and its MRE within wild-type vs. mutant NF2 −3′UTR (green). Dual-Luciferase assay of (I) HCT116 and (J) A549 cells co-transfected with wild-type or miR-92a-3p MRE mutant NF2 −3′UTR and miR-92a expression vectors. All experiments were performed in cells maintained in 0.5% serum. Data presented are representative of three independent trials and expressed as the mean ± standard deviation (SD). *P≤0.05, **P≤0.01, ***P≤0.001. NF2 , neurofibromin 2; UTR, untranslated region; MRE, microRNA response element.
Article Snippet: QuantiGene Singleplex miRNA probes were obtained from the pre-designed probes offered by eBioscience (Thermo Fisher Scientific, Inc.).
Techniques: Clone Assay, Sequencing, Luciferase, Transfection, Plasmid Preparation, Expressing, Construct, Mutagenesis, Standard Deviation