Mutant frequencies and spectra depend on growth state and passage number in cells cultured from transgenic lacZ-plasmid reporter mice

被引:23
作者
Busuttil, RA
Rubio, M
Dollé, MET
Campisi, J
Vijg, J
机构
[1] Univ Texas, Hlth Sci Ctr, Sam & Ann Barshop Inst Longev & Aging Studies, San Antonio, TX 78245 USA
[2] Univ Calif Berkeley, Lawrence Berkeley Lab, Div Life Sci, Berkeley, CA 94720 USA
[3] Natl Inst Publ Hlth & Environm, NL-3720 BA Bilthoven, Netherlands
[4] Buck Inst Age Res, Novato, CA USA
[5] S Texas Vet Hlth Care Syst, Ctr Geriatr Res Educ & Clin, San Antonio, TX USA
关键词
lacZ-plasmid mouse embryonic fibroblasts; ultraviolet (UV); mutation; quiescence; proliferation;
D O I
10.1016/j.dnarep.2005.07.006
中图分类号
Q3 [遗传学];
学科分类号
071007 ; 090102 ;
摘要
Transgenic mice harboring the lacZ gene within a plasmid that can be recovered and amplified in Escherichia coli, to establish mutant frequencies and spectra, have provided crucial insights into the relationships between mutations, cancer and aging in vivo. Here, we use embryonic fibroblasts from transgenic lacZ-plasmid reporter mice to determine the relationship between cell proliferation in culture and mutations induced by ultraviolet (UV) light. A single dose of 2.5 J/m(2) of UVC to actively proliferating cells caused an approximately eightfold increase in mutant frequency 24 h after irradiation. Identically treated quiescent cells showed a two-fold increase in mutant frequency. Thus, whereas proliferation facilitated the acquisition of mutations, it was not an absolute requirement. Characterization of the UV-induced mutations indicated that the lower mutant frequency in quiescent cells was due mainly to a reduction in point mutations; size-change mutations, indicative of translocations or deletions, were relatively unaffected by the growth state of the cells. To investigate long-term genomic stability after UVC-induced damage, we monitored the lacZ locus in irradiated cells passaged for many generations in culture. The results indicated the emergence of jackpot mutations of rapidly changing frequency, most likely reflecting the successive emergence and decline of dominant cell clones during long-term culture. These findings show that the lacZ-plasmid locus is a valid reporter for studying induced mutations in short-term cultures of both quiescent and proliferating fibroblasts. In long-term cultures, the locus is less suitable for studying induced mutations owing to the instability of the cell population. (C) 2005 Elsevier B.V. All rights reserved.
引用
收藏
页码:52 / 60
页数:9
相关论文
共 28 条
  • [1] Dynamics of DNA double-strand breaks revealed by clustering of damaged chromosome domains
    Aten, JA
    Stap, J
    Krawczyk, PM
    van Oven, CH
    Hoebe, RA
    Essers, J
    Kanaar, R
    [J]. SCIENCE, 2004, 303 (5654) : 92 - 95
  • [2] Proliferation is necessary for both repair and mutation in transgenic mouse cells
    Bielas, JH
    Heddle, JA
    [J]. PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA, 2000, 97 (21) : 11391 - 11396
  • [3] Quiescent murine cells lack global genomic repair but are proficient in transcription-coupled repair
    Bielas, JH
    Heddle, JA
    [J]. DNA REPAIR, 2004, 3 (07) : 711 - 717
  • [4] PLASMID-BASED TRANSGENIC MOUSE MODEL FOR STUDYING IN-VIVO MUTATIONS
    BOERRIGTER, METI
    DOLLE, MET
    MARTUS, HJ
    GOSSEN, JA
    VIJG, J
    [J]. NATURE, 1995, 377 (6550) : 657 - 659
  • [5] Oxygen accelerates the accumulation of mutations during the senescence and immortalization of murine cells in culture
    Busuttil, RA
    Rubio, M
    Dollé, MET
    Campisi, J
    Vijg, J
    [J]. AGING CELL, 2003, 2 (06) : 287 - 294
  • [6] Induction of cyclooxygenase-2 mRNA and protein expression in human gingival fibroblasts stimulated with nicotine
    Chang, YC
    Tsai, CH
    Yang, SH
    Liu, CM
    Chou, MY
    [J]. JOURNAL OF PERIODONTAL RESEARCH, 2003, 38 (05) : 496 - 501
  • [7] BIOCHEMICAL-ANALYSIS OF UV MUTAGENESIS IN ESCHERICHIA-COLI BY USING A CELL-FREE REACTION COUPLED TO A BIOASSAY - IDENTIFICATION OF A DNA REPAIR-DEPENDENT, REPLICATION-INDEPENDENT PATHWAY
    COHENFIX, O
    LIVNEH, Z
    [J]. PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA, 1992, 89 (08) : 3300 - 3304
  • [8] Evaluation of a plasmid-based transgenic mouse model for detecting in vivo mutations
    Dolle, MET
    Martus, HJ
    Gossen, JA
    Boerrigter, METI
    Vijg, J
    [J]. MUTAGENESIS, 1996, 11 (01) : 111 - 118
  • [9] Rapid accumulation of genome rearrangements in liver but not in brain of old mice
    Dolle, MET
    Giese, H
    Hopkins, CL
    Martus, HJ
    Hausdorff, JM
    Vijg, J
    [J]. NATURE GENETICS, 1997, 17 (04) : 431 - 434
  • [10] Dollé MET, 1999, ENVIRON MOL MUTAGEN, V34, P112, DOI 10.1002/(SICI)1098-2280(1999)34:2/3&lt