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Sh Alizadeh, S Kaviani, M Soleimani, Aa Pourfathollah, N Amirizadeh, F Kouhkan, S Abroun, M Noruzinia,
Volume 4, Issue 3 (20 2011)
Abstract

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.


Seyed Amir Hossein Emami, Shahin Mohammadi, Saeed Kavyani, Masud Soleimani, Shaban Alizadeh, Ehteram Dejbakhsh, Fatemeh Kouhkan, Majid Mossahebi, Hossein Dargahi,
Volume 5, Issue 3 (4 2011)
Abstract

Background and Aim: Micro RNAs are a group of small non-coding RNAs which play an important role in multiple processes such as proliferation, differentiation, apoptosis, and cancer. Recent studies indicate that mir-210 is overexpressed into erythroid linage during the differentiation of hematopoietic precursor. The main goal of the present study is to investigate the influence of mir-210 on the pattern of expression in hemoglobin gamma chain.

Materials and Methods: First, K562 cell line was cultured in RPMI1640 media. Then, pre-miR-210 was transferred into K562 cell line by lipofectamin. Finally, the alterations in the pattern of gamma chain expression were analyzed in days 7 and 14 by RT-PCR and real time PCR technique.

Results: It was demonstrated that the overexpression of mir-210 in K562 cell line would lead to a 25-fold increase in the expression of gamma chain in comparison with the control group. Data analysis revealed that the change in the pattern of hemoglobin gamma chain expression was meaningful (p<0.002).

Conclusion: Based on these data, overexpression of mir-210 can lead to a significant increase in the production of gamma chain. Therefore, more studies in the field may reveal the fact that an increase in mir-210 can be a suitable goal in the improvement of sickle cell anemia and β-thalassemia.



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