Journal of

Background and Aim: MicroRNAs (miRNA) are small noncoding RNA molecules that transcribed by RNA polymerase II. After biogenesis, these molecules act by incorporation into the RNA-induced silencing complex (RISC). MiRNAs are involved in multiple physiological and pathological processes such as proliferation, differentiation, apoptosis and cancer.
Recently several studies reported down regulation of mir-150 during erythropoesis. Since hemoglobin expression is valuable indicator of erythroid differentiation we evaluated the mir-150 downregulation effect on alpha chain expression by Quantitative RT-PCR.

Materials and Methods: K562cells were grown in RPMI1640 in standard condition. K562 cells were transfected by microRNA 150 Inhibitor using transfection kit .Mir-150 downregulation was confirmed by miRNA Real time PCR, followed by Q-RT-PCR to investigate the alpha chain expression changes.

Results: By relative QRT-PCR the alpha chain expression was increased 10 folds in comparison to untransfected and scramble cells. Furthermore, the differences were statistically significant (P<0.05)

Conclusion: Elevation of alpha chain expression in our study showed that mir-150 downregulation has a crucial role in erythroid differentiation and can introduce as a novel marker in alpha thalassemia. Further researches to find out the detail mechanism and miRNAs genes target could improve our knowledge about miRNAs potential in management of diseases and their applications in gene therapy and regenerative medicine.

 Background and Aim: Understanding the molecular mechanisms involved in the increased levels of HbF inducing drugs should be advised for effective induction. The aim of this study was to investigate the molecular effects of the drugs thalidomide and sodium butyrate considered as HbF inducer agents.

 Materials and Methods: In this experimental study, CD133+ cord blood stem cells carrying mutations of heterozygous β-thalassemia were isolated and differentiated into erythroid lineage. In order to evaluate the expression of the erythroid markers, CD71 and CD235a, was analysed. For this purpose, the RNA extracted from erythroid precursors at days 6 and 12 of erythroid differentiation and cDNA synthesized, and then the expression of these genes was performed by quantitative Real-time PCR technique.

 Results: The results of this study showed the significant effect of thalidomide on erythroid proliferation as compared to sodium butyrate and control group (P<0.05). Also, thalidomide significantly increased CD71expression and decreased CD235a expression as compared to sodium butyrate and control groups (P<0.05).

 Conclusion : Thalidomide may play its role on HbF induction by increasing the proliferation of early erythroid precursors.

Background and Aim: Different regulation processes have an effect on osteoblastic differentiation of mesenchymal stem cells (MSCs), and among them Wnt signaling pathway is particularly desirable. In Wnt signaling pathway, Adenomatous Polyposis Coli (APC) bind to β-catenin and induce its degradation, thereby acting as a negative regulator of canonical Wnt pathway. In this study, gene expression and DNA methylation of APC gene during osteoblastic differentiation were determined.

Materials and Methods: In this experimental study, after the isolation of MSCs, the induction of osteoblastic differentiation was done. To confirm osteoblastic differentiation, alizarin red staining together with the expression of Alkaline Phosphatase (ALP) and osteocalcin as specific osteoblastic markers was performed. APC gene methylation status by MSP (Methylation Specific PCR) and gene expression status of APC gene using Real-Time PCR technique during different times were evaluated.

Results: The results of alizarin red staining and the expression of ALP and osteocalcin confirmed osteoblastic differentiation. In addition, the results showed a significant decrease in the expression of APC gene on the 7^th day of osteoblastic differentiation (P<0.05). Also, the results revealed hypermethylation status of APC gene promoter during osteoblastic differentiation.

Conclusion: It seems that the decreased expression APC gene will play an important role in Wnt signaling pathway regulation in different stages during osteoblastic differentiation of bone marrow-derived MSC. Also, according to the results, APC gene promoter methylation will play an important role in controlling gene expression.