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

Abstract

in β-thalassemia, excessive α-globin chain impedes the normal development of red blood cells resulting in anemia. numerous mirnas, including mir-6747-3p, are aberrantly expressed in β-thalassemia major (β-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 β-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 β-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 γ-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 γ-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 β-thalassemia, highlighting mir-6747-3p as a potential molecular target for β-thalassemia therapy.