Differential induction of chromosomal instability by DNA strand-breaking agents.

To investigate the role of DNA strand breakage as the molecular lesion responsible for initiating genomic instability, five different strand-breaking agents, bleomycin, neocarzinostatin, hydrogen peroxide, restriction endonucleases, and ionizing radiation, were examined for their capacity to induce delayed chromosomal instability. These studies used GM10115 human-hamster hybrid cells, which contain one copy of human chromosome 4 in a background of 20-24 hamster chromosomes. Chromosomal instability was investigated using fluorescence in situ hybridization to visualize chromosomal rearrangements involving the human chromosome. Rearrangements are detected multiple generations after treatment, in clonal populations derived from single progenitor cells surviving treatment of the specified DNA-damaging agents. Clastogenic and cytotoxic activities of all agents were tested by examining chromosome aberration yields in first-division metaphases and by clonogenic survival assays. Analysis of over 250 individual clones representing over 50,000 metaphases demonstrates that when compared at comparable levels of cell kill, ionizing radiation, bleomycin, and neocarzinostatin are equally effective at eliciting delayed genomic instability. These observations document, for the first time, the persistent destabilization of chromosomes following chemical treatment. In contrast, the analysis of nearly 300 clones and 60,000 metaphases, involving treatment with four different restriction endonucleases and/or hydrogen peroxide, did not show any delayed chromosomal instability. These data indicate that DNA strand breakage per se does not necessarily lead to chromosomal instability but that the complexity or quality of DNA strand breaks are important in initiating this phenotype.

[1]  C. Streffer,et al.  Do DNA double-strand breaks induced by Alu I lead to development of novel aberrations in the second and third post-treatment mitoses? , 1996, Radiation research.

[2]  I. Goldberg.  Free radical mechanisms in neocarzinostatin-induced DNA damage. , 1987, Free radical biology & medicine.

[3]  W. Morgan,et al.  X-ray induction of methotrexate resistance due todhfr gene amplification , 1990, Somatic cell and molecular genetics.

[4]  M. Siciliano,et al.  Preferential loss or inactivation of chromosome Z4APRT allele in CHO cells , 1989, Somatic cell and molecular genetics.

[5]  J. Little,et al.  Evidence that DNA double-strand breaks initiate the phenotype of delayed reproductive death in Chinese hamster ovary cells. , 1992, Radiation research.

[6]  D T Goodhead,et al.  Alpha-particle-induced chromosomal instability in human bone marrow cells. , 1994, Lancet.

[7]  A. Johansson,et al.  Clonal chromosome aberrations and genomic instability in X-irradiated human T-lymphocyte cultures. , 1993, Mutation research.

[8]  J. Ward.  Radiation mutagenesis: the initial DNA lesions responsible. , 1995, Radiation research.

[9]  J. Liesch,et al.  Identification of thymidine-5'-aldehyde at DNA strand breaks induced by neocarzinostatin chromophore. , 1982, Proceedings of the National Academy of Sciences of the United States of America.

[10]  Shirley A. Miller,et al.  A simple salting out procedure for extracting DNA from human nucleated cells. , 1988, Nucleic acids research.

[11]  J. Stubbe,et al.  Mechanisms of bleomycin-induced DNA degradation , 1987 .

[12]  D. Vanderwall,et al.  Sequence-specific isotope effects on the cleavage of DNA by bleomycin. , 1989, Science.

[13]  G. Adair,et al.  Mutagenicity testing in mammalian cells. I. Derivation of a Chinese hamster ovary cell line heterozygous for the adenine phosphoribosyltransferase and thymidine kinase loci. , 1980, Mutation research.

[14]  B. Dutrillaux,et al.  Chromosomal instability and alterations of telomeric repeats in irradiated human fibroblasts. , 1994, International journal of radiation biology.

[15]  J. Phillips,et al.  Illegitimate recombination induced by DNA double-strand breaks in a mammalian chromosome , 1994, Molecular and cellular biology.

[16]  J. Day,et al.  Delayed chromosomal instability induced by DNA damage , 1993, Molecular and cellular biology.

[17]  J. Day,et al.  Genomic instability induced by ionizing radiation. , 1996, Radiation research.

[18]  M. Siciliano,et al.  Spontaneous CHO APRT heterozygotes reflect high-frequency, allele-specific deletion of the chromosome Z4 APRT gene. , 1989, Somatic cell and molecular genetics.

[19]  R. Ullrich,et al.  Latent expression of p53 mutations and radiation-induced mammary cancer. , 1995, Cancer research.

[20]  A. Goddard,et al.  Chromosome aberrations and cancer. , 1991, Science.

[21]  C. Limoli,et al.  Radiation vs Chemical Damage to DNA , 1987 .

[22]  J. Ward,et al.  DNA damage produced by ionizing radiation in mammalian cells: identities, mechanisms of formation, and reparability. , 1988, Progress in nucleic acid research and molecular biology.

[23]  J. Ward,et al.  Mammalian cells are not killed by DNA single-strand breaks caused by hydroxyl radicals from hydrogen peroxide. , 1985, Radiation research.

[24]  B. Dutrillaux,et al.  Specific chromosome instability induced by heavy ions: a step towards transformation of human fibroblasts? , 1993, Mutation research.

[25]  L. Loeb,et al.  Mutator phenotype may be required for multistage carcinogenesis. , 1991, Cancer research.

[26]  M. Bender,et al.  Current status of cytogenetic procedures to detect and quantify previous exposures to radiation. , 1988, Mutation research.

[27]  W. Bradley,et al.  High-frequency nonrandom mutational event at the adenine phosphoribosyltransferase (aprt) locus of sib-selected CHO variants heterozygous foraprt , 1982, Somatic cell genetics.

[28]  P. Nowell.  The clonal evolution of tumor cell populations. , 1976, Science.

[29]  D. T. Goodhead,et al.  Transmission of chromosomal instability after plutonium α-particle irradiation , 1992, Nature.

[30]  K. Brookman,et al.  Validation of conditions for efficient detection of HPRT and APRT mutations in suspension-cultured Chinese hamster ovary cells. , 1980, Mutation research.

[31]  C. Limoli,et al.  Radiation and hydrogen peroxide induced free radical damage to DNA. , 1987, The British journal of cancer. Supplement.

[32]  R. Stallings,et al.  High-frequency structural gene deletion as the basis for functional hemizygosity of the adenine phosphoribosyltransferase locus in Chinese hamster ovary cells. , 1983, Proceedings of the National Academy of Sciences of the United States of America.

[33]  M. Siciliano,et al.  Chromosomal rearrangements and gene expression in CHO Cells: Mapping of alleles for eight enzyme loci on CHO chromosomes Z3, Z4, Z5, and Z7 , 1984, Somatic cell and molecular genetics.

[34]  I. Goldberg,et al.  3'-Formyl phosphate-ended DNA: high-energy intermediate in antibiotic-induced DNA sugar damage. , 1987, Proceedings of the National Academy of Sciences of the United States of America.

[35]  M. Bauchinger,et al.  A proposed system for scoring structural aberrations detected by chromosome painting. , 1995, Cytogenetics and cell genetics.