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Determination of γ-globin induction levels by different concentrations of hydroxyurea in K562 cell lines for β thalassemia treatment
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Zahra Deilami Khiabani   1, Mahdi Banan , Ali Mohammad Asgharian , Jalal Gharesouran , Gholamreza Javadi , Kimia Kahrizi , Hossein Najmabadi |
| 1- , zdeilami@yahoo.com |
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Abstract: (20572 Views) |
Background: Hydroxyurea (HU) is currently used for β thalassemia treatment through induction of fetal γ-globin. In this study, effects of different concentrations of hydroxyurea on induction of γ-globin gene in K562 cells, and inhibitory effect of siRNA against candidate gene which may be involved in HU mediated γ-globin induction were assessed.
Materials and Methods: In this experimental study, K562 cells were treated by different concentrations of HU (0, 50, 100 and 200 μM). siRNA against candidate gene in HU mediated γ-globin gene induction was transfected to K562 cells using lipofectamine 2000. The level of γ-globin gene induction and inhibition were determined by quantitative real-time PCR.
Results: There were 1.75-, 2.45- and 2.55- fold inductions in γ- globin gene using 50, 100 and 200μM HU, respectively. There has been 79.5% down regulation on candidate gene in siRNA transfected K562 cells.
Conclusion: This study showed that γ-globin induction in 100μM HU is similar to 200 μM HU treated K562 cells. There was also efficient inhibitory effect on candidate gene which may be involved in HU mediated γ-globin induction. |
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| Keywords: γ-globin gene, Hydroxyurea, K562 cells, Real time PCR, siRNA |
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Full-Text [PDF 1153 kb]
(3445 Downloads)
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Subject:
Epidemiology
Received: 2006/09/6 | Published: 2009/10/15
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