PMID- 14977355
OWN - NLM
STAT- MEDLINE
DCOM- 20040903
LR  - 20191108
IS  - 0965-0407 (Print)
IS  - 0965-0407 (Linking)
VI  - 14
IP  - 4-5
DP  - 2004
TI  - Hydroxyurea-induced apoptosis in an EBV-immortalized lymphoblastoid cell line.
PG  - 235-45
AB  - Hydroxyurea (HU) is an inhibitor of nucleotide synthesis extensively used to 
      control the chronic phase of myeloid leukemia. This antimetabolite has been 
      employed in the clinic for several decades but in recent years the leukemogenic 
      potential of HU has been suspected. In the present study, a B-lymphoblastoid cell 
      line transformed by the Epstein-Barr virus was used to investigate the apoptotic 
      effects of HU and delineate some of the molecular pathways implicated in the 
      cytotoxic action. The cell line, characterized by immunophenotyping, cytogenetic 
      and fluorescence in situ hybridization (FISH) studies, showed no chromosomal 
      abnormalities, even after a prolonged exposure to HU. Different flow cytometry 
      assays were used to measure HU-induced impairment of the cell cycle, inhibition 
      of DNA synthesis, and the occurrence of apoptosis. The treatment with HU leads to 
      the appearance of a hypo-diploid DNA content peak (sub-G1) characteristic of the 
      apoptotic cell population. The drug also induces a cell block in S phase as 
      measured by 5-bromo-2'-deoxyuridine (BrdU) incorporation. Inhibition of DNA 
      synthesis precedes induction of apoptosis by HU. A drug-induced loss of plasma 
      membrane asymmetry was characterized by flow cytometry using annexin V-FITC to 
      stain phosphatidylserine residues. The implication of the antiapoptotic protein 
      Bcl-2 and the tumor suppressor p53 in the development of HU-mediated apoptosis 
      was also evidenced. The drug appears to promote cell death by regulating the 
      expression levels of these two proteins. Different criteria define the apoptotic 
      response of the lymphoblastoid cells to the treatment with HU. However, the 
      extent of drug-induced cell death is limited, and no DNA fragmentation and no 
      activation of the caspase cascade was observed in this model. Beyond the specific 
      interest in HU-induced apoptosis, the work reported here illustrates the utility 
      of the EBV immortalization process to investigate the pharmacological activity of 
      specific drugs from clinical samples.
FAU - Huyghe, Pauline
AU  - Huyghe P
AD  - INSERM U-524, Institut de Recherches sur le Cancer de Lille, 1 Place de Verdun, 
      59045 Lille cedex, France.
FAU - Dassonneville, Laurent
AU  - Dassonneville L
FAU - Fenaux, Pierre
AU  - Fenaux P
FAU - Bailly, Christian
AU  - Bailly C
LA  - eng
PT  - Journal Article
PT  - Research Support, Non-U.S. Gov't
PL  - United States
TA  - Oncol Res
JT  - Oncology research
JID - 9208097
RN  - 0 (Proto-Oncogene Proteins c-bcl-2)
RN  - 0 (Tumor Suppressor Protein p53)
RN  - G34N38R2N1 (Bromodeoxyuridine)
RN  - X6Q56QN5QC (Hydroxyurea)
SB  - IM
MH  - Apoptosis/*drug effects
MH  - Bromodeoxyuridine/metabolism
MH  - Cell Cycle/drug effects
MH  - Cell Line, Transformed
MH  - Cell Membrane/metabolism
MH  - Flow Cytometry
MH  - Herpesvirus 4, Human/*physiology
MH  - Humans
MH  - Hydroxyurea/*pharmacology
MH  - Immunophenotyping
MH  - In Situ Hybridization, Fluorescence
MH  - Leukemia, Myelogenous, Chronic, BCR-ABL Positive/metabolism/*pathology/*virology
MH  - Proto-Oncogene Proteins c-bcl-2/metabolism
MH  - Tumor Cells, Cultured
MH  - Tumor Suppressor Protein p53/metabolism
EDAT- 2004/02/24 05:00
MHDA- 2004/09/04 05:00
CRDT- 2004/02/24 05:00
PHST- 2004/02/24 05:00 [pubmed]
PHST- 2004/09/04 05:00 [medline]
PHST- 2004/02/24 05:00 [entrez]
AID - 10.3727/000000003772505362 [doi]
PST - ppublish
SO  - Oncol Res. 2004;14(4-5):235-45. doi: 10.3727/000000003772505362.