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Evaluation of the biological role of Peroxiredoxins 1, 2 and 6 on erythrocyte cells by RNA silencing

Grant number: 13/09427-7
Support Opportunities:Scholarships in Brazil - Master
Effective date (Start): October 01, 2013
Effective date (End): May 31, 2015
Field of knowledge:Biological Sciences - Biochemistry - Molecular Biology
Principal Investigator:Anderson Ferreira da Cunha
Grantee:Carla Peres de Paula
Host Institution: Centro de Ciências Biológicas e da Saúde (CCBS). Universidade Federal de São Carlos (UFSCAR). São Carlos , SP, Brazil


Reactive oxygen species (ROS) are products generated by normal metabolism, exposure to external agents or as a secondary response to various diseases. At low physiological levels ROS act as messengers "redox" in intracellular signaling and regulation, while in excess can cause various damages to the cells. To combat this damage, cells have developed enzymatic and non-enzymatic systems in different cellular compartments to maintain an adequate level of ROS and regulate its action. Among the enzymatic mechanisms peroxiredoxins (PRDXs) stand out for their great abundance and reactivity with their substrates. In humans have been described six isoforms of PRDXs localized in different cellular compartments, which have important functions in various cell signaling cascades, and play an important role as an antioxidant, protecting the cell from oxidative damage. It was demonstrated that in mammalian cells exposed to high levels of molecular oxygen, such as erythrocytes, the PRDX are very abundant. Furthermore, studies have shown that modulation of the level of ROS has a close relationship with hemolytic anemias such as glucose-6-phosphate dehydrogenase (G6PD) deficiency, sickle cell anemia and alpha and beta thalassemia. Under these conditions, the oxidative damage caused by hemolysis of the cells favor the formation of such species which further aggravates the patient's condition. Therefore, this study aims to assess the biological role played by peroxiredoxins PRDX1, PRDX2 PRDX3 in erythrocytes using the technique of gene silencing of each protein using RNA interference in K562 and KU812 cells. Thus, we aim to evaluate in response to the silencing of each PRDX, the cell viability, intracellular levels of ROS and changes in the level of expression of other PRDX. We believe that understanding the molecular mechanisms involved in protection against ROS in erythrocytes can collaborate with a better understanding of the pathophysiology of hemolytic anemias, identifying possible targets to help in the management and to reduce the effects of the disease in these patients.

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