Introduction: Thalassemias are among the most common autosomal recessive monogenic diseases, which occur due to mutations in the beta-globin gene and lead to a decrease or non-synthesis of the globin chain and thus a reduction in hemoglobin levels. Induced production of the γ -globin chains reduces α/β-chain imbalance in β-thalassemia through the creation of fetal Hb (HbF), thereby increasing effective erythropoiesis and reducing hemolysis and the level of anemia. A score of studies has identified that HU is well accepted in clinical practice and can be associated as another option for β-thalassemia treatment. the exact mechanisms by which HU induces HbF production are not fully understood and are still controversial. we applied Weighted Gene Co-expression Network Analysis (WGCNA) approach to identify and quantify changes in gene expression that were reflected in the HU-treated human erythroblastic leukemia cells.
Methods: LIN28B and CA1 were selected as two related genes in the switching and expression of fetal hemoglobin and their expression behavior was evaluated under HU treatment. K562 cell line was cultured and cells for examination in two groups of control and treatment with hydroxyurea in 3 concentrations of 50,100 and 150 μM and 24, 48, and 72 hours with 3 replications were cultured. The RNA was extracted by RNA Extraction Kit Then, cDNA synthesis was conducted and quantitative PCR (qRT-PCR) was accomplished.
Results: LIN28B showed an increased expression level in all treated groups compared to the controls. But CA1 expression showed a decrease in all-time series and dosage of treatment. Along with this γ-globin gene expression was significantly elevated.
Conclusion: The LIN28 gene is known to regulate the let-7 family of miRNAs, and the expression of LIN28 transcripts is associated with the inhibition of let-7. It has been shown that the expression of LIN28 protein in adult erythrocytes not only leads to a decrease of let-7 miRNAs expression but also can upregulate HbF expression. Since the CA1 gene is involved in the c-MYB factor pathway, its downregulation under HU treatment conditions suggests the key role of the c-MYB pathway in HbF expression. C-MYB is a key factor in regulating HbF production and may be involved in the modification of the globin gene by controlling cell cycles.