The identification of molecules useful for pharmacologically-mediated regulation of the expression of human gamma-globin genes is a crucial step for the development of agents potentially useful in experimental therapy of hematological disorders, including beta-thalassemia and sickle cell anemia. It is well established, indeed, that increase of fetal hemoglobin (HbF) to 30% of the total hemoglobin (hB) leads to a significant improvement of the clinical status of patients affected by these hematological disorders. In this respect, a rapid and reproducible analysis of the effects of potential HbF inducers on accumulation of mRNA for gamma-, beta- and alpha-globins is relevant. Here, we review data on the use of real-time reaction (RT-PCR) for studying accumulation of mRNAs for gamma-, beta- and alpha-globins. By this procedure it is possible to differentiate inducers able to stimulate preferential expression of gamma-globin genes from inducers able to stimulate both gamma-globin and beta-globin mRNA production. This analytical strategy is of potential clinical significance to identify HbF inducers useful to alleviate the symptoms underlying beta-thalassemia and sickle cell anemia, and compounds stimulating beta-globin gene expression, of interest in a variety of beta+-thalassemias.
Quantitative RT-PCR for the analysis of expression of alpha-, beta- and gamma-globin genes in erythroid cells
BIANCHI, Nicoletta;BORGATTI, Monica;GAMBARI, Roberto
2003
Abstract
The identification of molecules useful for pharmacologically-mediated regulation of the expression of human gamma-globin genes is a crucial step for the development of agents potentially useful in experimental therapy of hematological disorders, including beta-thalassemia and sickle cell anemia. It is well established, indeed, that increase of fetal hemoglobin (HbF) to 30% of the total hemoglobin (hB) leads to a significant improvement of the clinical status of patients affected by these hematological disorders. In this respect, a rapid and reproducible analysis of the effects of potential HbF inducers on accumulation of mRNA for gamma-, beta- and alpha-globins is relevant. Here, we review data on the use of real-time reaction (RT-PCR) for studying accumulation of mRNAs for gamma-, beta- and alpha-globins. By this procedure it is possible to differentiate inducers able to stimulate preferential expression of gamma-globin genes from inducers able to stimulate both gamma-globin and beta-globin mRNA production. This analytical strategy is of potential clinical significance to identify HbF inducers useful to alleviate the symptoms underlying beta-thalassemia and sickle cell anemia, and compounds stimulating beta-globin gene expression, of interest in a variety of beta+-thalassemias.I documenti in SFERA sono protetti da copyright e tutti i diritti sono riservati, salvo diversa indicazione.
