Efeito do tratamento com hidroxiureia sobre a expressão gênica de ATF4, MSN, PBXIPI e SNCA em pacientes com anemia falciforme e em células K562

Abstract

Our study aimed to analyze the expression and protein content of four genes (ATF4, MSN, PBXIP1 and SNCA) by qPCR and Western Blotting in individuals with sickle cell anemia treated and not treated with hydroxyurea (HU) in order to establish associations with pathophysiology sickle cell anemia and, mainly, with HU treatment. To prove the results, analyzes were also performed with K562 cells culture stimulated for erythroid differentiation in the presence and absence of HU. Our results show decrease in the expression of MSN in individuals with the disease, which can be justified by the increase in cell adhesion in this group. The data obtained also suggest that treatment with HU induces the expression of PBXIPI and inhibits SNCA. The PBXIPI activation would assist in stimulating the differentiation and proliferation of hematopoietic cells, while the decrease in the content of SNCA may be associated with the reduction of inflammatory processes, which is an effect of great importance in worsening the disease. For the ATF4 gene, associated with oxidative stress and neovascularization, we expected that treatment with hydroxyurea would lead to a reduction in expression, however, we did not obtain a statistically significant difference in the comparison between groups. Finally, we suggest that the increase in oxidative stress by adding H2O2 to the culture medium also influences the expression of our genes and affects the potential effect of hydroxyurea.