Increased repair of γ-induced DNA double-strand breaks at lower dose-rate in CHO cells

被引:15
|
作者
Boucher, D [1 ]
Hindo, J [1 ]
Averbeck, D [1 ]
机构
[1] Ctr Univ Orsay, Inst Curie, Sect Rech, CNRS,UMR 2027,IC,CEA, F-91405 Orsay, France
关键词
gamma-irradiation; Chinese hamster ovary cells (CHO); nonhomologous end-joining (NHEJ); low dose-rate; deferoxamine;
D O I
10.1139/Y04-006
中图分类号
R9 [药学];
学科分类号
1007 ;
摘要
DNA double-strand breaks (DSBs) are highly cell damaging. We asked whether for a given dose a longer irradiation time would be advantageous for the repair of DSBs: Varying the gamma-irradiation dose and its delivery time (0.05 Gy/min low dose-rate (LDR) compared with 3.5 Gy/min high dose-rate), confluent Chinese hamster ovary cells (CHO-K1) and Ku80 mutant cells (xrs-6) deficient in nonhomologous end-joining (NHEJ) were irradiated in agarose plugs at room temperature using a cesium-137 gamma-ray source. We used pulsed-field gel electrophoresis (PFGE) to measure DSBs in terms of the fraction of activity released (FAR). At LDR, one third of DSBs were repaired in CHO-K1 but not in xrs-6 cells, indicating the involvement of NHEJ in the repair of gamma-induced DSBs at:a prolonged irradiation incubation time. To improve DSB measurements, we introduced in our PFGE protocol an antioxidant at the cell lysis step, thus avoiding free-radical, side reactions on DNA and spurious DSBs. Addition of the metal-chelator deferoxamine (DFO) decreased more efficiently the basal DSB level than did reduced glutathione (GSH), showing that measuring DSBs in the absence of DFO reduces precision and underestimates the role of NHEJ in the dose-rate effect on DSB yield.
引用
收藏
页码:125 / 132
页数:8
相关论文
共 50 条
  • [31] Dose-rate effect on radiation-induced DNA double-strand breaks in the human fibroblast HF19 cell line
    Foray, N
    Fertil, B
    Alsbeih, MGA
    Badie, C
    Chavaudra, N
    Iliakis, G
    Malaise, EP
    INTERNATIONAL JOURNAL OF RADIATION BIOLOGY, 1996, 69 (02) : 241 - 249
  • [32] Gamma-irradiated quiescent cells repair directly induced double-strand breaks but accumulate persistent double-strand breaks during subsequent DNA replication
    Minakawa, Yusuke
    Atsumi, Yuko
    Shinohara, Akira
    Murakami, Yasufumi
    Yoshioka, Ken-ichi
    GENES TO CELLS, 2016, 21 (07) : 789 - 797
  • [33] The Influence of Bromodeoxyuridine on the Induction and Repair of DNA Double-Strand Breaks in Glioblastoma Cells
    Nadine N. Nusser
    Detlef Bartkowiak
    Erwin M. Röttinger
    Strahlentherapie und Onkologie, 2002, 178 : 504 - 509
  • [34] Identification of genes involved in repair of DNA double-strand breaks in mammalian cells
    Jeggo, PA
    RADIATION RESEARCH, 1998, 150 (05) : S80 - S91
  • [35] Efficient repair of DNA double-strand breaks in malignant cells with structural instability
    Cheng, Yue
    Zhang, Zhenhua
    Keenan, Bridget
    Roschke, Anna V.
    Nakahara, Kenneth
    Aplan, Peter D.
    MUTATION RESEARCH-FUNDAMENTAL AND MOLECULAR MECHANISMS OF MUTAGENESIS, 2010, 683 (1-2) : 115 - 122
  • [36] INDUCTION AND REPAIR OF DOUBLE-STRAND BREAKS IN THE REPLICATING DNA OF HELA-CELLS
    WARTERS, RL
    LYONS, BW
    RADIATION RESEARCH, 1993, 134 (03) : 337 - 342
  • [37] The influence of bromodeoxyuridine on the induction and repair of DNA double-strand breaks in glioblastoma cells
    Nusser, NN
    Bartkowiak, D
    Röttinger, EM
    STRAHLENTHERAPIE UND ONKOLOGIE, 2002, 178 (09) : 504 - 509
  • [38] DIFFERENCES IN REPAIR OF DOUBLE-STRAND BREAKS IN HUMAN-TUMOR CELL-LINES DURING LOW DOSE-RATE IRRADIATION
    CASSONI, AM
    MCMILLAN, TJ
    PEACOCK, JH
    STEEL, GG
    INTERNATIONAL JOURNAL OF RADIATION BIOLOGY, 1989, 56 (05) : 843 - 844
  • [39] INDUCTION AND REPAIR OF SINGLE-STRAND AND DOUBLE-STRAND BREAKS IN DNA IN ATAXIA TELANGIECTASIA CELLS
    VANDERSCHANS, GP
    CENTEN, HB
    JASPERS, NGJ
    REGULSKI, MR
    VANZEELAND, AA
    MUTATION RESEARCH, 1982, 96 (01): : 149 - 150
  • [40] Sources of DNA Double-Strand Breaks and Models of Recombinational DNA Repair
    Mehta, Anuja
    Haber, James E.
    COLD SPRING HARBOR PERSPECTIVES IN BIOLOGY, 2014, 6 (09):