MiR-20b directly targets 3’-UTR to regulate the expression of HIF-1α

Abstract:

Objective：To investigate whether miRNA-20b（miRNA-20b/miR-20b） may bind to the 3’-untranslated region（3’-UTR） of hypoxia inducible factor-1α（HIF-1α） mRNA to regulate the expression of HIF-1α. Methods：Targetscan and miRanda softwares were used to predict the presence or absence of miR-20b binding site in the HIF-1α 3’-UTR region. The dual luciferase gene re－porter assay system（pmiR-RB-ReportTM-HIF-1α） was constructed and identified. The ratios of dual luciferase activity were detected in HeLa cells 24 hours after mimics-miR-20b（M） or NC-miR-20b（N） and recombinant plasmid（RP） co-transfection. This study was divided into six groups：RP，RP+M，RP+N，P，P+M and P+N groups. Results：There was a miR-20b binding site in the 3’-UTR of HIF-1α mRNA. The double luciferase gene reporter vector containing HIF-1α 3’-UTR was successfully constructed. The dual lu－ciferase activity was significantly lower in recombinant plasmid and mimics-miR-20b co-transfection group than in empty group（P=0.022）. Conclusion：miR-20b can directly bind to HIF-1α 3’-UTR region to regulate the expression of HIF-1α.