Journal: Oncology Letters
Article Title: MicroRNA-23a/27a/24-2 cluster promotes gastric cancer cell proliferation synergistically
doi: 10.3892/ol.2018.8924
Figure Lengend Snippet: SOCS6 is a target gene of miR-27a and miR-24. (A) The putative binding sites of miR-23a, miR-27a and miR-24 on SOCS6 were predicted using Targetscan. The interspecies sequence alignment revealed the mature miRNA sequences and the putative binding sites. Luciferase reporter constructs containing WT or mutant target sites of the 3′-untraslated region of SOCS6 mRNA for miR-23a, miR-27a and miR-24, respectively. (B) SC-M1 cells were co-transfected with dual luciferase reporter plasmid, containing a specific miRNA-binding sequence (wild-type or mutant), and 90 nM miR inhibitor for 24 h. Dual-luciferase activities were measured using a microplate reader. *P<0.05 and **P<0.01 for mutant vs. wild-type. (C) SC-M1 cells were treated with miR-23a, miR-27a and miR-24 inhibitors mixture for 48 h. The expression of SOCS6 was examined by western blotting. SOCS6, suppressor of cytokine-induced signaling 6; miR, microRNA; WT, wild-type.
Article Snippet: MicroRNA analyses were commissioned by the High-throughput Genome Analysis Core Facility of the VYM Genome Research Center (National Yang-Ming University, Taipei, Taiwan) on NCode TM Multi-Species miRNA microarrays (Invitrogen; Thermo Fisher Scientific, Inc.).
Techniques: Binding Assay, Sequencing, Luciferase, Construct, Mutagenesis, Transfection, Plasmid Preparation, Expressing, Western Blot