The Targeted Regulating Role of Has-miR-577 and Has-miR-583 on Gene FGF-21 Detected by the Dual

Objective To determine the targeted regulating role of has-miR-577 and has-miR-583 on the expression of fibroblast growth factor 21 (FGF-21) based on a constructed luciferase reporter FGF-21 gene vector. Methods The site of has-miR-577 and has-miR-583 target genes FGF-21 were predicted by the bioinformatics analyzing tools online．FGF-21 gene fragments, combined with has-miR-577 or has-miR-583 sequences and mutant sequences,were designed and synthesized.The wild type (psiCHECK2-FGF-21) and mutant (psiCHECK2-FGF-21-mut) luciferase reporter gene carriers were constructed．The relevant plasmids 〔hsa-miR-577mimics, hsa-miR-583 mimics or miR negative control (miR-NC)〕 and luciferase reporter gene carrier (wild type or mutant ) were co-transfected into 293T cells．The luciferase reporter system was used to detect the luciferase activity．The effects of has-miR-577 and has-miR-583 on the expression of FGF-21 were observed. Results The double enzyme electrophoresis and sequencing results showed that the gene fragment size and sequences of the wild type (psiCHECK2-FGF-21) and mutant (psiCHECK2-FGF-21-mut) carriers met expectations of the experiment. The luciferase assays revealed that has-miR-577 and has-miR-583 significantly diminished luciferase activity from the reporter vector containing 3′UTR of FGF-21 (PFGF-21-mut)．Conclusion FGF-21 gene can be targeted by has-miR-577 and has-miR-583.