Upregulation of miR-6747-3p affects red blood cell lineage development and induces fetal hemoglobin expression by targeting BCL11A in β-thalassemia

In beta-thalassemia, excessive alpha-globin chain impedes the normal development of red blood cells resulting in anemia. Numerous miRNAs, including miR-6747-3p, are aberrantly expressed in beta-thalassemia major (beta-TM), but there are no reports on the mechanism of miR-6747-3p in regulating red blood cell lineage development and fetal hemoglobin (HbF) expression. In the present study, RT-qPCR was utilized to confirm miR-6747-3p expression in patients with beta-TM and the healthy controls. Electrotransfection was employed to introduce the miR-6747-3p mimic and inhibitor in both HUDEP-2 and K562 cells, and red blood cell lineage development was evaluated by CCK-8 assay, flow cytometry, Wright-Giemsa staining and Benzidine blue staining. B-cell lymphoma/leukemia 11A (BCL11A) was selected as a candidate target gene of miR-6747-3p for further validation through FISH assay, dual luciferase assay and Western blotting. The results indicated that miR-6747-3p expression was notably higher in patients with beta-TM compared with healthy controls and was positively related to HbF levels. Functionally, miR-6747-3p overexpression resulted in the hindrance of cell proliferation, promotion of cell apoptosis, facilitation of cellular erythroid differentiation and gamma-globin expression in HUDEP-2 and K562 cells. Mechanistically, miR-6747-3p could specifically bind to the 546-552 loci of BCL11A 3 '-UTR and induce gamma-globin expression. These data indicate that upregulation of miR-6747-3p affects red blood cell lineage development and induces HbF expression by targeting BCL11A in beta-thalassemia, highlighting miR-6747-3p as a potential molecular target for beta-thalassemia therapy.