The Effect of miR-940 Up-regultion on HbF and Erythroid Markers Expression in k-562 Cell Line

Abstract

Abstract Background: Fetal hemoglobin (α2γ2) is the main oxygen transport protein in the human fetus. Fetal hemoglobin is nearly completely replaced by hemoglobin A, except in a few thalassemia cases and sickle cell anemia. Several studies have indicated that expression of γ-globin might be regulated post-transcriptionally. Small non-coding RNA called microRNAs which target mRNA can lead to translated repression or mRNA decay. The aim of this study is to investigate the effect of miR-940 up-regultion on γ-chain gene expression and erythroid markers in k562 cell line. Materials and Methods: In this experimental study, k562 cells were cultured in RPMI1640. Then pre miR-940 was transfected by electroporation method in k562 cell line. In 3, 7 and 14 days, RNA was extracted and cDNA synthesized in selected days. Up-regulation of miR-940 was confirmed by miRNA Quantitative real time PCR and then the expression γ of chains and GATA-1 was investigated by QRT-PCR. Finally, erythroid markers were checked by flow cytometry. Results: In 3, 7 and 14 days after transfection, the GATA-1 and γ-chain expression were increased in comparison with untransfected cells. Also, the expression of erythroid markers was increased. Conclusion: The data show that up-regulation of miR-940 has a role in the increase of γ-chain gene expression in k-562 cell line. We suggest that miR-940 may be a significant potent therapeutic target for increasing Hb F level. Patients with sickle cell anemia and β-thalassemia are suitable candidate for treatment in this way