Induction of gamma-globin mRNA, erythroid differentiation and apoptosis in UVA-irradiated human erythroid cells in the presence of furocumarin derivatives.
Psoralens, also known as furocoumarins, are a class of photosensitizers largely used in the therapy of various skin diseases. In this study we have evaluated the combined effects of UVA irradiation and furocoumarins derivatives on (a) erythroid differentiation and apoptosis of human leukemia K562 cells and (b) globin gene expression in cultures of human erythroid progenitors derived from the peripheral blood. To prove the activity of a series of linear and angular furocoumarins derivatives, we employed the human leukemia K562 cell line and the two-phase liquid culture procedure for growing erythroid progenitors. Quantitative real-time reverse transcription polymerase-chain assay (Q-RT-PCR) was employed for quantification of the accumulation of globin mRNAs. The results obtained demonstrate that both linear and angular furocoumarins are strong inducers of erythroid differentiation of K562 cells. From a preliminary screening, we have selected two derivatives, 5-methoxypsoralen (5-MOP) and trimethylangelicin (TMA), for which we have investigated their mechanism of action. The cell cycle analysis showed that these derivatives induce, after irradiation, a cell cycle arrest in the G2/M phase, followed by apoptosis. Mitochondrial depolarisation and caspases activation seem to be involved in the mechanism of cell death. In erythroid precursor cells, psoralens in combination with UVA irradiation, stimulate at very low concentrations a preferential increase of gamma-globin mRNA. Altogether, these data suggest that psoralen derivatives warrant further evaluation as potential therapeutic drugs in beta-thalassemia and sickle cell anemia.
Viola
Giampietro
G
Department of Pharmaceutical Sciences, University of Padova, Via Marzolo 5, 35131 Padova, Italy. giampietro.viola.1@unipd.it
Vedaldi
Daniela
D
Dall'Acqua
Francesco
F
Fortunato
Elena
E
Basso
Giuseppe
G
Bianchi
Nicoletta
N
Zuccato
Cristina
C
Borgatti
Monica
M
Lampronti
Ilaria
I
Gambari
Roberto
R
D017209:Apoptosis / Q000187:drug effects / Q000235:genetics / Q000528:radiation effects; D002453:Cell Cycle / Q000187:drug effects / Q000528:radiation effects; D002454:Cell Differentiation / Q000187:drug effects / Q000235:genetics / Q000528:radiation effects; D004249:DNA Damage; D041905:Erythroid Cells / Q000187:drug effects / Q000378:metabolism / Q000473:pathology / Q000528:radiation effects; D005434:Flow Cytometry; D011564:Furocoumarins / Q000737:chemistry / Q000494:pharmacology*; D015870:Gene Expression / Q000187:drug effects; D020869:Gene Expression Profiling; D005914:Globins / Q000096:biosynthesis*; D006801:Humans; D020014:K562 Cells; D053078:Membrane Potential, Mitochondrial / Q000187:drug effects / Q000235:genetics / Q000528:radiation effects; D010718:Phosphatidylserines / Q000096:biosynthesis; D012333:RNA, Messenger / Q000096:biosynthesis*; D017382:Reactive Oxygen Species / Q000378:metabolism; D020133:Reverse Transcriptase Polymerase Chain Reaction; D014466:Ultraviolet Rays
D011564:Furocoumarins; D010718:Phosphatidylserines; D012333:RNA, Messenger; D017382:Reactive Oxygen Species; D005914:Globins