Hydroxyurea-induced apoptosis in an EBV-immortalized lymphoblastoid cell line

被引:12
作者
Huyghe, P
Dassonneville, L
Fenaux, P
Bailly, C
机构
[1] Inst Rech Canc Lille, INSERM U524, F-59045 Lille, France
[2] CHU Lille, Hematol Lab, Serv Malad Sang, F-59045 Lille, France
关键词
hydroxyurea; apoptosis; cell cycle; p53; Bcl-2;
D O I
10.3727/000000003772505362
中图分类号
R73 [肿瘤学];
学科分类号
100214 ;
摘要
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-G(1)) 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.
引用
收藏
页码:235 / 245
页数:11
相关论文
共 37 条
[1]   INDUCTION OF APOPTOSIS IN CHRONIC MYELOGENOUS LEUKEMIA LYMPHOCYTES BY HYDROXYUREA AND ADRIAMYCIN [J].
ANAND, S ;
VERMA, H ;
KUMAR, L ;
SINGH, N .
CANCER LETTERS, 1995, 88 (01) :101-105
[2]   Sickle cell anaemia - Progress in pathogenesis and treatment [J].
Ballas, SK .
DRUGS, 2002, 62 (08) :1143-1172
[3]   Effect of antimetabolite drugs of nucleotide metabolism on the anti-human immunodeficiency virus activity of nucleoside reverse transcriptase inhibitors [J].
Balzarini, J .
PHARMACOLOGY & THERAPEUTICS, 2000, 87 (2-3) :175-187
[4]  
BENE MC, 1995, LEUKEMIA, V9, P1783
[5]  
Boersma AWM, 1996, CYTOMETRY, V24, P123, DOI 10.1002/(SICI)1097-0320(19960601)24:2<123::AID-CYTO4>3.0.CO
[6]  
2-K
[7]   Gene expression profiling in chronic myeloid leukemia patients treated with hydroxyurea [J].
Bruchova, H ;
Borovanova, T ;
Klamova, H ;
Brdicka, R .
LEUKEMIA & LYMPHOMA, 2002, 43 (06) :1289-1295
[8]  
DACIE JV, 1984, PRACTICAL HAEMATOLOG, P50
[9]   Successful treatment of chronic myeloid leukemia during pregnancy with hydroxyurea [J].
Fadilah, SAW ;
Ahmad-Zailani, H ;
Soon-Keng, C ;
Norlaila, M .
LEUKEMIA, 2002, 16 (06) :1202-1203
[10]   The role of phosphatidylserine in recognition of apoptotic cells by phagocytes [J].
Fadok, VA ;
Bratton, DL ;
Frasch, SC ;
Warner, ML ;
Henson, PM .
CELL DEATH AND DIFFERENTIATION, 1998, 5 (07) :551-562