The Analysis and Identification of MiR-27 Target Gene

MicroRNAs ( miRNAs) is singlestranded oligoribonucleotides about 20 - 25 nucleotides ( nt) in length and function at many biological process,for example,cell proliferation,apoptosis,growth,differentiation and metabolism. MiR-27 play an important role in adipogenesis and muscle stem cell behavior,but it is unclear about its role in the development of neural cell. In the present study,the target genes of miR-27 was analyzed by miRBase and TargetScan databases. A recombinant miR-27 eukaryotic expression vector was constructed and expressed it in human glioma cells U343. We reconstructed the firefly luciferase and renilla luciferase vector and fused the 3′-UTR of miR-27 target gene Bmi1 with report gene that was transfected in U343 cells. luciferase activity was detected by Dual-Luciferase Reporter ( DLR) Assay. Results showed the common target genes of miR-27a and miR-27b regulated mainly the process of development. MiR-27 eukaryotic expression vector can produce mature miR-27a and miR-27b. Firefly luciferase activity was significantly reduced by cotransfection of miR-27a (P 0. 05),miR-27b (P 0. 05),combination of both (P 0. 01) and sense 3′-UTR of Bmi1. These results implied that Bmi1 may be the one of miR-27 target genes.