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2007 | 54 | 2 | 281-287
Article title

Gamma-radiation-induced ATM-dependent signalling in human T-lymphocyte leukemic cells, MOLT-4

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Abstracts
EN
ATM kinase (ATM) is essential for activation of cell cycle check points and DNA repair in response to ionizing radiation (IR). In this work we studied the molecular mechanisms regulating DNA repair and cell death in human T-lymphocyte leukemic cells, MOLT-4. Apoptosis was evaluated by flow-cytometric detection of annexin V. Early apoptotic cells were determined as sub-G1 cells and late apoptotic cells were determined as APO2.7-positive ones. Proteins involved in ATM signalling pathway were analysed by Western-blotting. We observed a rapid (0.5 h) phosphorylation of ATM declining after 6 h after irradiation by all the doses studied (1.5, 3.0, and 7.5 Gy). Checkpoint kinase-2 (Chk-2) was also phosphorylated after 0.5 h but its phosphorylated form persisted 4, 2, and 1 h after the doses of 1.5, 3.0, and 7.5 Gy, respectively. The amount of p53 protein and its form phosphorylated on Ser-392 increased 1 h after irradiation (1-10 Gy). The lethal dose of 7.5 Gy caused an immediate induction and phosphorylation of p53 after 0.5 h post-irradiation. At the time of phosphorylation of p53, we found simultaneous phosphorylation of the oncoprotein Mdm2 on Ser-166. Neither ATM nor its downstream targets showed a dose-dependent response after 1 h when irradiated by the doses of 1-10 Gy. MOLT-4 cells were very sensitive to the effect of IR. Even low doses, such as 1.5 Gy, induced apoptosis 16 h after irradiation (evaluated according to the cleavage of nuclear lamin B to a 48-kDa fragment). IR-induced molecular signalling after exposure to all the tested doses was triggered by rapid phosphorylation of ATM and Chk-2. Subsequent induction of p53 protein and its phosphorylation was accompanied by concomitant phosphorylation of its negative regulator, oncoprotein Mdm2, and followed by induction of apoptosis.
Publisher

Year
Volume
54
Issue
2
Pages
281-287
Physical description
Dates
published
2007
received
2007-01-16
revised
2007-05-21
accepted
2007-06-01
(unknown)
2007-06-12
Contributors
author
  • Department of Radiobiology, Faculty of Military Health Sciences in Hradec Králové, University of Defence in Brno, Brno, Czech Republic
  • Institute of Medical Biochemistry, Faculty of Medicine in Hradec Králové, Charles University in Prague, Prague, Czech Republic
  • Institute of Medical Biochemistry, Faculty of Medicine in Hradec Králové, Charles University in Prague, Prague, Czech Republic
  • Department of Radiobiology, Faculty of Military Health Sciences in Hradec Králové, University of Defence in Brno, Brno, Czech Republic
  • Institute of Clinical Immunology and Allergology, University Hospital Hradec Králové, Hradec Králové, Czech Republic
  • Department of Radiobiology, Faculty of Military Health Sciences in Hradec Králové, University of Defence in Brno, Brno, Czech Republic
  • Department of Radiobiology, Faculty of Military Health Sciences in Hradec Králové, University of Defence in Brno, Brno, Czech Republic
  • Institute of Medical Biochemistry, Faculty of Medicine in Hradec Králové, Charles University in Prague, Prague, Czech Republic
  • Department of Radiobiology, Faculty of Military Health Sciences in Hradec Králové, University of Defence in Brno, Brno, Czech Republic
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Document Type
Publication order reference
Identifiers
YADDA identifier
bwmeta1.element.bwnjournal-article-abpv54p281kz
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