Induction of gamma-globin gene transcription by hydroxycarbamide in primary erythroid cell cultures from Lepore patients.
Increased expression of fetal haemoglobin (HbF) may ameliorate the clinical course of beta-thalassemia and sickle cell disease. Some pharmacological agents, such as hydroxycarbamide (HC), can increase fetal haemoglobin synthesis during adult life. Cellular selection and/or molecular mechanisms have been proposed to account for this increase. To explore the mechanism of action of HC we focused on homozygous Hb-Lepore patients that presented with high fetal haemoglobin levels and were good responders to HC treatment "in vivo". We performed primary erythroid cultures from peripheral blood of four homozygous Lepore patients. The increase in HBG (gamma-globin) transcription levels and HbF content in these cultures, after HC treatment, were detected by quantitative real time polymerase chain reaction analysis and flow cytometric analysis. Primary transcript "in-situ" hybridization analysis showed a 2-fold increase in the number of cells expressing both HBG alleles in HC-treated erythroid cultures. These studies, demonstrating the larger number of biallelic HBG expressing cells, suggest that HC is able to stimulate the activation of HBG transcription. These observations provide evidences that the molecular mechanism of action is involved in the increase of fetal haemoglobin production by HC.
Calzolari
Roberta
R
Unità di Ricerca "P. Cutino", Ematologia II, Università degli Studi di Palermo, Palermo, Italy.
Pecoraro
Alice
A
Borruso
Vito
V
Troia
Antonio
A
Acuto
Santina
S
Maggio
Aurelio
A
Di Marzo
Rosalba
R
D002478:Cells, Cultured; D041905:Erythroid Cells / Q000187:drug effects*; D005319:Fetal Hemoglobin / Q000096:biosynthesis; D005914:Globins / Q000096:biosynthesis* / Q000235:genetics; D006455:Hemoglobins, Abnormal / Q000235:genetics*; D006801:Humans; D006918:Hydroxyurea / Q000494:pharmacology*; D017404:In Situ Hybridization, Fluorescence; D012333:RNA, Messenger / Q000235:genetics; D014158:Transcription, Genetic / Q000187:drug effects; D017086:beta-Thalassemia / Q000097:blood* / Q000235:genetics
D006455:Hemoglobins, Abnormal; D012333:RNA, Messenger; D005914:Globins; D005319:Fetal Hemoglobin; C037659:hemoglobin Lepore; D006918:Hydroxyurea