High Frequency Of Chromatid Breaks Research Articles

Bleomycin-induced chromosomal damage and shortening of telomeres in peripheral blood lymphocytes of incident cancer patients.

Disruption of genomic integrity due to deficient DNA repair capacity and telomere shortening constitute hallmarks of malignant diseases. Incomplete or deficient repair of DNA double-strand breaks (DSB) is manifested by chromosomal aberrations and their frequency reflects inter-individual differences of response to exposure to mutagenic compounds. In this study, we investigated chromosomal integrity in peripheral blood lymphocytes (PBL) from newly diagnosed cancer patients, including 47 breast (BC) and 44 colorectal cancer (CRC) patients and 90 matched healthy controls. Mutagen sensitivity was evaluated by measuring chromatid breaks (CTAs) induced by bleomycin and supplemented by the chemiluminescent measurement of γ-H2AX phosphorylation in 19 cancer patients (11 BC, 8 CRC). Relative telomere length (RTL) was determined in 22 BC, 32 CRC, and 64 controls. We observed statistically significant increased level of CTAs (P=.03) and increased percentage of aberrant cells (ACs) with CTAs (P=.05) in CRC patients compared with controls after bleomycin treatment. No differences were observed between BC cases and corresponding controls. CRC and BC patients with shorter RTL (below median) exhibited significantly higher amount of ACs (P=.02), CTAs (P=.02), and cells with high frequency of CTAs (≥12 CTAs/PBL; P=.03) after bleomycin treatment. No such associations were observed in healthy controls. γ-H2AX phosphorylation after bleomycin treatment in PBL did not differ between CRC and BC patients. Our results suggest that altered DSB repair measured by sensitivity towards mutagen in PBL occurs particularly in CRC carcinogenesis. Irrespective of cancer type, telomere shortening may be associated with a decreased capacity to repair DSB.

Association between TP53 gene ARG72PRO polymorphism and chromosome aberrations in human cancers

It is well known that the TP53 gene considerably influences on DNA repair processes. Polymorphisms in the TP53 gene, particularly the well-known Arg72Pro in codon 72 of exon 4 (Ex4+119 G>C; rs1042522), can modify the functionality of the p53 protein and activation of DNA repair. Actually, polymorphic variants Arg and Pro were found to have different properties of regulation of TP53-dependent DNA repair target genes, that can effect various levels of chromosome aberrations in cancer patients with these genotypes. Here, we studied frequency of chromatid breaks (CB), chromosome-type aberrations (CTA) and aberrant cells (AC) in cancer patients (n = 102) with various Arg72Pro genotypes. It was shown that the Arg variant of TP53 gene is associated with high frequency of AC and chromatid breaks. That is Arg/Arg carriers have more different chromosome aberrations in comparison to individuals with Arg/Pro and Pro/Pro genotypes (P < 0.05). Conversely, the lowest level of AC and chromatid breaks were detected in cancer patients with the Pro/Pro genotype. A completely unexpected result was that women with Arg/Arg genotype had the most high frequency of CB and AC in comparison to Arg/Pro and Pro/Pro women carriers (P < 0.001). In the group of male patients we did not show any differences in chromosome aberrations between carriers of Arg72Pro genotypes. In conclusion, the TP53 gene Arg72Pro polymorphism appreciably influence on occurrence of chromosome aberrations in cancer.

G2 chromatid damage and repair kinetics in normal human fibroblast cells exposed to low- or high-LET radiation

Radiation-induced chromosome damage can be measured in interphase using the Premature Chromosome Condensation (PCC) technique. With the introduction of a new PCC technique using the potent phosphatase inhibitor calyculin-A, chromosomes can be condensed within five minutes, and it is now possible to examine the early damage induced by radiation. Using this method, it has been shown that high-LET radiation induces a higher frequency of chromatid breaks and a much higher frequency of isochromatid breaks than low-LET radiation. The kinetics of chromatid break rejoining consists of two exponential components representing a rapid and a slow time constant, which appears to be similar for low- and high-LET radiations. However, after high-LET radiation exposures, the rejoining process for isochromatid breaks influences the repair kinetics of chromatid-type breaks, and this plays an important role in the assessment of chromatid break rejoining in the G2 phase of the cell cycle.

Alterações cromossômicas causadas pela radiação dos monitores de vídeo de computadores

Concerns were raised about the potential damaging effects of electromagnetic field (EMF) radiation emissions of ELF (extremely low frequency) and VLF (very low frequency) computer video display monitors (VDM), it was assessed the frequency of structural chromosome abnormalities and investigated the cell cycle kinetics in individuals occupationally exposed to VDM radiation. Chromosome aberrations were investigated in 2,000 first cell cycle metaphases obtained after 48-hour cultures of peripheral blood lymphocytes drawn from 10 individuals occupationally exposed to VDM radiation (group E) and 10 controls (group C). Cell cycle kinetics was studied using the mitotic index (MI) and cellular proliferation index (CPI). Statistical analysis showed significantly higher frequencies of anomalous metaphases (E=5.9%; C=3.7%) and anomaly/cell (E=0.066+/-0.026; C= 0.040+/-0.026) in individuals exposed to VDM radiation. The most common cytogenetic alterations seen were chromatid breaks at frequencies of 0.034+/-0.016 in group E and 0.016+/-0.015 in group C. There was no significant difference between MI and CPI frequencies in both groups. The study findings suggest genotoxic effects of EMF emissions revealed by the higher frequency of chromatid breaks in individuals exposed to VDM radiation. However, there is a need of further studies on EMF genetic effects using other investigation methods.

Gamma-radiation sensitivity and risk of glioma.

About 9% of human cancers are brain tumors, of which 90% are gliomas. gamma-Radiation has been identified as a risk factor for brain tumors. In a previous pilot study, we found that lymphocytes from patients with glioma were more sensitive to gamma-radiation than were lymphocytes from matched control subjects. In this larger case-control study, we compared the gamma-radiation sensitivity of lymphocytes from glioma patients with those from control subjects and investigated the association between mutagen sensitivity and the risk for developing glioma. We used a mutagen sensitivity assay (an indirect measure of DNA repair activity) to assess chromosomal damage. We gamma-irradiated (1.5 Gy) short-term lymphocyte cultures from 219 case patients with glioma and from 238 healthy control subjects frequency matched by age and sex. After irradiation, cells were cultured for 4 hours, and then Colcemid was added for 1 hour to arrest cells in mitosis. Fifty metaphases were randomly selected for each sample and scored for chromatid breaks. All statistical tests were two-sided. We observed a statistically significantly higher frequency of chromatid breaks per cell from case patients with glioma (mean = 0.55; 95% confidence interval [CI] = 0.50 to 0.59) than from control subjects (mean = 0.44; 95% CI = 0.41 to 0.48) (P<.001). Using 0.40 (the median number of chromatid breaks per cell in control subjects) as the cut point for defining mutagen sensitivity and adjusting for age, sex, and smoking status, we found that mutagen sensitivity was statistically significantly associated with an increased risk for glioma (odds ratio = 2.09; 95% CI = 1.43 to 3.06). When the data were divided into tertiles, the relative risk for glioma increased from the lowest tertile to the highest tertile (trend test, P<.001). gamma-Radiation-induced mutagen sensitivity of lymphocytes may be associated with an increased risk for glioma, a result that supports our earlier preliminary findings.

Suppression of X-Ray-Induced Chromatid Breaks in Human Tumor Cells by Introduction of Normal Chromosome 4

Human tumor cells in culture frequently show an infinite lifespan and are characterized by an abnormally high frequency of chromatid breaks after x-irradiation during G 2 phase of the cell cycle. This abnormally high frequency of persistent chromatid breaks results from deficient repair of the radiation-induced DNA damage. In three of four tumor cell lines, addition of a single human chromosome 4 from normal cells by microcell fusion resulted in efficient repair as evidenced by suppression of radiation-induced chromatid breaks to the level in normal cells. With respect to senescence-related gene (s), two of these four tumor cell lines belonged to complementation group A and one each to groups C and D. Chromosome 4 restored DNA repair efficiency in only one of the two tumor cell lines of complementation group A. These results suggest that chromosome 4 carries a DNA repair gene or gene(s) that complement the repair deficiency in three of these four tumor lines. Furthermore, the gene(s) involved in cellular senescence on chromosome 4 appear to differ from the gene(s) for repair of radiation-induced DNA damage during G 2.

Radiation–Induced Chromatid Breaks and DNA Repair in Blood Lymphocytes of Patients with Dysplastic Nevi and/or Cutaneous Melanoma

Each chromatid contains a single continuous molecule of double-stranded DNA, so chromatid breaks represent unrepaired DNA double-strand breaks. Frequencies of chromatid breaks after G2 phase x-irradiation were determined in phytohemagglutinin-stimulated blood lymphocytes from normal subjects and from four categories of patients with dysplastic nevi with or without cutaneous melanoma or with melanoma alone. Some cells were treated with an inhibitor of DNA repair replication to determine enzymatic incision activity at damaged sites after exposure to x-rays, UVC, or fluorescent light. Whereas one of 16 normal controls had deficient DNA repair, all 17 patients from families with hereditary dysplastic nevi with or without melanoma (category I) had a deficiency in repair of radiation-induced DNA damage, manifested as an abnormally high frequency of chromatid breaks after x-irradiation or a reduced capacity to incise the damaged sites after UV exposure. Four of 11 patients with sporadic dysplastic nevi alone (category II) and eight of 12 with sporadic dysplastic nevi and melanoma (category III) showed deficient DNA repair after x-irradiation. One of two patients with sporadic melanoma and no dysplastic nevi (category IV) was also deficient in repair of x-ray-induced damage. Deficient DNA repair thus appears to be associated with hereditary dysplastic nevi with or without melanoma. It also characterizes some patients with sporadic dysplastic nevi or melanoma.

Cytogenetic Response to G 2 -Phase X Irradiation in Relation to DNA Repair and Radiosensitivity in a Cancer-Prone Family with Li-Fraumeni Syndrome

Noncancerous skin fibroblasts from six family members with Li-Fraumeni syndrome, five with cancer of diverse tissue origin and one with a premalignant neoplasm, showed a high frequency of chromatid aberrations, 94 to 119 breaks and 58 to 95 gaps per 100 metaphase cells arrested with colcemid 0.5 to 1.5 h after X irradiation (1.75 x 10(-2) C/kg). This response results from deficient repair of the radiation-induced DNA damage. In contrast, skin fibroblasts from two unrelated normal controls and a spouse showed 19 breaks and 17 to 19 gaps per 100 cells. Whereas all six members of the cancer-prone family had a radioresistant phenotype, only four had an inherited p53 mutation. Fibroblasts from a radioresistant family member showed the same extent of chromatid damage directly (0 to 0.5 h) after G2-phase X irradiation as those from the radiosensitive control spouse. We conclude, therefore, that radiosensitivity, as determined by cell killing in asynchronous populations of skin fibroblasts, is unrelated to chromosomal sensitivity to G2-phase X irradiation. However, the persistence of a high frequency of chromatid breaks and gaps at 0.5 to 1.5 h after G2-phase X irradiation, a manifestation of deficient DNA repair, is associated with proneness to cancer in this family.

X-ray-induced chromatid damage in relation to DNA repair and cancer incidence in family members.

The cytogenetic response to G2-phase X-irradiation was examined in phytohemagglutinin-stimulated peripheral-blood lymphocytes from 69 individuals, a few of whom were cancer patients. The cancer patients had not received radiation or chemotherapy. The responses of cells arrested by Colcemid 30 to 90 min after X-irradiation (58R) could be divided into 2 distinct categories: 51 individuals had aberration frequencies typical of normal individuals in previous studies, while 18 others had a 2- to 3-fold higher frequency of chromatid breaks and gaps. Because chromatid breaks and gaps result from unrepaired DNA strand breaks, the first category may represent an efficient DNA repair phenotype, while the second may represent a deficient repair phenotype. The individuals with the deficient G2 response reported having first- and second-degree relatives with a 3.6- and 2.2-fold higher mean frequency of cancer, respectively. The present results, together with those from earlier studies of families with a genetic disorder predisposing to cancer, suggest that this deficient cytogenetic response to G2 phase X-irradiation is associated with a high risk of cancer.

INSITU NICK TRANSLATION, AN ALTERNATIVE TO CYTOGENETICS IN ANALYZING MUTAGEN EFFECTS ON HUMAN-CELLS

A chromosomal break indicates a double-stranded DNA break of the chromatin fiber. Cytogenetic observations can enumerate chromosome breaks but cannot detect single-stranded DNA breaks (nicks). When a high frequency of chromosomal breaks occurs in a cell population, whether spontaneously occurred or mutagen induced, a higher frequency of nicks is expected. In order to correlate these two type of DNA lesions, we used human lymphoblastoid lines known to be either hypersensitive or relatively resistant to the genotoxicity of bleomycin. The frequency of nicks was estimated by grain counts over interphase nuclei in autoradiographs following the technique of in situ nick translation. Good agreement was found between these two sets of data, viz, hypersensitive lines showed a high frequency of chromatid breaks as well as a high frequency of grain counts, and vice versa. These results suggest that the phenomenon of differential bleomycin sensitivity is genetically determined.

Role of DNA repair in malignant neoplastic transformation of human mammary epithelial cells in culture.

Epithelial cells derived from normal human mammary tissue were examined for capacity to repair radiation-induced chromatin DNA damage. Repair capacity was estimated by quantifying chromatid aberrations in metaphase cells arrested 0.5-1.5 h after X-irradiation during G2. The parental cells at passage 12 had 19 chromatid breaks and 16 gaps per 100 metaphase cells, representing efficient repair. Of two continuous cell lines, derived after benzo[a]pyrene treatment, A1 maintained the efficient repair phenotype through passage 50, while a subline of A1 developed the repair-deficient phenotype characterized by a 3- to 5-fold higher frequency of chromatid breaks or gaps. This line was transformed to tumorigenic cells by HaMSV and SV40 T antigen. The second continuous line B5 and derivatives had 102-165 chromatid breaks and 87-134 gaps per 100 metaphases (deficient repair phenotype). This line was transformed to tumorigenic cells by KiMSV. As reported previously for human epidermal keratinocytes, acquisition of this repair-deficient phenotype appears to be an early requisite step in the malignant neoplastic transformation of human cells in culture.

Carrier detection in xeroderma pigmentosum.

We were able to detect clinically normal carriers of xeroderma pigmentosum (XP) genes with coded samples of either peripheral blood lymphocytes or skin fibroblasts, using a cytogenetic assay shown previously to detect individuals with cancer-prone genetic disorders. Metaphase cells of phytohemagglutinin-stimulated T-lymphocytes from eight individuals who are obligate heterozygotes for XP were compared with those from nine normal controls at 1.3, 2.3, and 3.3 h after x-irradiation (58 R) during the G2 phase of the cell cycle. Lymphocytes from the XP heterozygotes had twofold higher frequencies of chromatid breaks or chromatid gaps than normal (P less than 10(-5)) when fixed at 2.3 or 3.3 h after irradiation. Lymphocytes from six XP homozygotes had frequencies of breaks and gaps threefold higher than normal. Skin fibroblasts from an additional obligate XP heterozygote, when fixed approximately 2 h after x-irradiation (68 R), had a twofold higher frequency of chromatid breaks and a fourfold higher frequency of gaps than fibroblasts from a normal control. This frequency of aberrations in cells from the XP heterozygote was approximately half that observed in the XP homozygote. The elevated frequencies of chromatid breaks and gaps after G2 phase x-irradiation may provide the basis of a test for identifying carriers of the XP gene(s) within known XP families.

Cytogenetical characterization of Chinese hamster ovary X-ray-sensitive mutant cells xrs 5 and xrs 6: III. Induction of cell killing, chromosomal aberrations and sister-chromatid exchanges by bleomycin, mono- and bi-functional alkylating agents

Two X-ray-sensitive mutants of CHO-K1 cells, xrs 5 and xrs 6, were characterised with regard to their responses to genotoxic chemicals, namely bleomycin, MMS, EMS, MMC and DEB for induction of cell killing, chromosomal aberrations and SCEs at different stages of the cell cycle. In addition, induction of mutations at the HPRT and Na +/K + ATPase (Oua) loci was evaluated after treatment with X-rays and MMS. Xrs 5 and xrs 6 cells were more sensitive than wild-type CHO-K1 to the cell killing effect of bleomycin (3 and 13 times respectively) and for induction of chromosomal aberrations (3 and 4.5 times). In these mutants a higher sensitivity for induction of chromosomal aberrations to MMS, EMS, MMC and DEB was observed (1.5–3.5 times). The mutants also showed increased sensitivity for cell killing effects of mono- and bi-functional alkylating agents (1.7–2.5 times). The high cell killing effect of X-rays in these mutants was accompanied by a slight increase in the frequency of HPRT mutation. The xrs mutants were also more sensitive to MMS for the increased frequency of TG r and Oua r mutants when compared to wild-type CHO-K1 cells. Though bleomycin is known to be a poor inducer of SCEs, an increase in the frequency of SCEs in xrs 6 cells (doubling at 1.2 μg/ml) was found in comparison to no significant increase in xrs 5 or CHO-K1 cells. The induced frequency of SCEs in all cell types increased in a similar way after the treatment with mono- or bi-functional alkylating agents. MMS treatment of G 2-phase cells yielded a higher frequency of chromatid breaks in the mutants in a dose-dependent manner compared to no effect in wild-type CHO-K1 cells. Treatment of synchronised mutant cells at G 1 stage with bleomycin resulted in both chromosome- and chromatid-type aberrations (similar to the response to X-ray treatment) in contrast to the induction of only chromosome-type aberrations in wild-type CHO-K1 cells. The frequency of chromosomal aberrations (chromosome and chromatid types) also increased with MMC treatment in G 1 cells of xrs mutants. DEB treatment of G 1 cells induced mainly chromatid-type aberrations in all cell types. The possible reasons for the increased sensitivity of xrs mutants to the chemical mutagens studied are discussed and the results are compared to cells derived from radiosensitive ataxia telangiectasia patients.

G 2 chromosomal radiosensitivity of ataxia-telangiectasia heterozygotes

Five lines of skin fibroblasts from individuals heterozygous for ataxia-telangiectasia (A-T), compared with six cell lines from age-matched normal controls, show a much higher frequency of chromatid breaks and gaps following x-irradiation during the G 2 phase of the cell cycle. The magnitude of this difference suggests that G 2 chromatid radiosensitivity could provide the basis for an assay to detect A-T heterozygotes. Though clinically normal, A-T heterozygotes share a high risk of cancer with A-T homozygotes and constitute approximately 1% of the human population. Further, we propose that G 2 chromosomal radiosensitivity, which appears to result from a DNA repair deficiency, may be associated with a genetic predisposition to cancer.

Clastogenicity of furans found in food

Cultured Chinese hamster ovary (CHO) cells were exposed for 3 h to furan and 6 furan derivatives (furfural, furfuryl alcohol, 5-methyl furfural, 2-methyl furan, 2,5-dimethyl furan and 2-furyl methyl ketone). Each of the 6 furan derivatives induced a relatively high frequency of chromatid breaks and chromatid exchanges in the absence of a liver microsomal activation preparation. The response of the furans to the addition of an S9 mixture differed. The clastogenic activities of 5-methyl furfural, 2-furyl methyl ketone, furfural and furfuryl alcohol were increased, whereas that of 2-methyl furan and 2,5-dimethyl furan were significantly decreased. Furan itself showed a clastogenic activity only in the presence of an S9 mixture.

N-nitrosomrpholine and N-nitrosobutylamine-stimulated DNA synthesis in BHK-21/C13 cells

Non-semiconservative DNA synthesis was examined in BHK-21/C13 cells after treatment with the aliphatic N-nitrosobutylamine (NB) and the heterocyclic N-nitrosomorpholine (NM). The extent of repair synthesis after alkylation was compared quentitatively and calculations were made of the amount of DNA damage per cell and number of bases inserted per damaged site. Of the chromosome aberrations caused by NM the most predominant were dicentrics. Other aberrations included fragments, gaps, breaks and exchanges of both the chromatid and chromosome type. Unlike NM, NB elicted a higher frequency of chromatid breaks.

High Frequency of Chromatid Breaks in Two In vitro Cell Populations

Previous article No AccessLetters to the EditorsHigh Frequency of Chromatid Breaks in Two In vitro Cell PopulationsT. C. Hsu and G. K. MannaT. C. Hsu Search for more articles by this author and G. K. Manna Search for more articles by this author PDFPDF PLUS Add to favoritesDownload CitationTrack CitationsPermissionsReprints Share onFacebookTwitterLinkedInRedditEmail SectionsMoreDetailsFiguresReferencesCited by The American Naturalist Volume 93, Number 870May - Jun., 1959 Published for The American Society of Naturalists Article DOIhttps://doi.org/10.1086/282077 Views: 4Total views on this site Citations: 13Citations are reported from Crossref PDF download Crossref reports the following articles citing this article:N. C. Popescu, B. C. Casto, J. A. Dipaolo Infrequent chromosome aberrations in syrian hamster cells following partial synchrony by amino-acid deprivation, Journal of Cellular Physiology 86, no.S2S2 (Dec 1975): 599–604.https://doi.org/10.1002/jcp.1040860505Arthur D. Bloom Induced Chromosomal Aberrations in Man, (Jan 1972): 99–172.https://doi.org/10.1007/978-1-4757-4429-3_3M.K. Patterson UPTAKE AND UTILIZATION OF AMINO ACIDS BY CELL CULTURES, (Jan 1972): 171–209.https://doi.org/10.1016/B978-0-12-598301-3.50012-XJ.J. Freed, Susan A. Schatz Chromosome aberrations in cultured cells deprived of single essential amino acids, Experimental Cell Research 55, no.33 (Jun 1969): 393–409.https://doi.org/10.1016/0014-4827(69)90574-6Carlo Valenti, Shama K. Vethamany Functional anatomy of a cytogenetic service, American Journal of Obstetrics and Gynecology 99, no.33 (Oct 1967): 434–457.https://doi.org/10.1016/S0002-9378(16)34558-6Samuel Irwin, Jose Egozcue Chromosomal Abnormalities in Leukocytes from LSD-25 Users, Science 157, no.37863786 (Jul 1967): 313–314.https://doi.org/10.1126/science.157.3786.313M. M. Cohen, M. J. Marinello, N. Back Chromosomal Damage in Human Leukocytes Induced by Lysergic Acid Diethylamide, Science 155, no.37683768 (Mar 1967): 1417–1419.https://doi.org/10.1126/science.155.3768.1417M. Fraccaro, Anna Mannini, L. Tiepolo, C. Zara High frequency of spontaneous recurrent chromosome breakage in an untreated human tumour, Mutation Research/Fundamental and Molecular Mechanisms of Mutagenesis 2, no.66 (Dec 1965): 559–561.https://doi.org/10.1016/0027-5107(65)90023-0 BIBLIOGRAPHY, (Jan 1965): 731–782.https://doi.org/10.1016/B978-0-08-010664-9.50023-4J.R. Gibbs, G.P. Arden SIMPLIFIED MANAGEMENT OF HEAD INJURIES, The Lancet 281, no.72797279 (Mar 1963): 505.https://doi.org/10.1016/S0140-6736(63)92411-5J. Wahrman, E. Robinson CHROMOSOME DAMAGE AFTER X-RAY THERAPY, The Lancet 281, no.72797279 (Mar 1963): 505–506.https://doi.org/10.1016/S0140-6736(63)92412-7T.C. Hsu Chromosomal Evolution in Cell Populations, (Jan 1962): 69–161.https://doi.org/10.1016/S0074-7696(08)60539-2 Hally J. Sax , and Kari N. Passano Spontaneous Chromosome Aberrations in Human Tissue Culture Cells, The American Naturalist 95, no.881881 (Oct 2015): 97–102.https://doi.org/10.1086/282163