cytochalasin-b has been researched along with Ataxia-Telangiectasia* in 4 studies
4 other study(ies) available for cytochalasin-b and Ataxia-Telangiectasia
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Use of the cytokinesis-block micronucleus assay to measure radiation-induced chromosome damage in lymphoblastoid cell lines.
To establish the optimal experimental conditions for the use of the micronuclei (MN) test to determine the level of chromosomal damage induced by ionising radiation (IR) exposure in lymphoblastoid cell lines, a time-course study was performed comparing a normal and an ataxia telangiectasia (AT) cell line, the latter being characterised by an extreme radiation sensitivity. Several parameters were analysed: the use of cytochalasin-B (Cyt-B) to quantify MN, the optimum fixation time to measure radiation-induced MN, the most appropriate treatment dose of IR to distinguish between the normal and the radiosensitive cells and the cell-cycle distribution after irradiation. The results obtained showed that the spontaneous as well as the radiation-induced levels of MN were significantly higher in the AT cell compared to the normal cells (P < 0.001 and P = 0.005, respectively). In both cell types the number of radiation-induced MN were lower in the cultures without Cyt-B than those with Cyt-B (P < 0.001), with the AT cells being distinguished in terms of IR-induced MN from the normal cells only with the addition of Cyt-B. The level of MN formation was independent of the dose of Cyt-B used (3 or 6 microg/ml). The optimum time for radiation-induced MN measured was found to be between 48 and 72 h post-irradiation, with 2 and 4 Gy exposures inducing similar levels of MN. However, as the higher dose caused a greater delay in the cell-cycle, treatment with 2 Gy with MN measurement at 48 h in the presence of 3 microg/ml Cyt-B were chosen as the optimum experimental conditions. This choice was validated using two additional normal and AT cell lines. In conclusion, our results show that the use of Cyt-B increases the sensitivity of the MN test for comparing differences in radiosensitivity between lymphoblastoid cell lines. Topics: Ataxia Telangiectasia; Ataxia Telangiectasia Mutated Proteins; Cell Cycle; Cell Cycle Proteins; Cell Division; Cell Line; Chromosome Aberrations; Chromosomes, Human; Cytochalasin B; DNA Damage; DNA-Binding Proteins; Humans; Lymphocytes; Micronucleus Tests; Protein Serine-Threonine Kinases; Radiation Tolerance; Tumor Suppressor Proteins | 2003 |
Micronucleus induction by neocarzinostatin and methyl methanesulfonate in ionizing radiation--sensitive Chinese hamster V79 cell mutants.
Two mutant V79 Chinese hamster cell lines (V-E5, XR-V15B) which show hypersensitivities to DNA damage and their two parental cell lines (V79-LE, V79-B) were used for micronucleus studies. The characteristics of V-E5 strongly resemble those of cells derived from patients suffering from the genomic instability syndrome ataxia telangiectasia, whereas XR-V15B has a decreased ability to rejoin double-strand breaks. The two cell lines V-E5 and XR-V15B showed increased spontaneous micronucleus frequencies and higher sensitivity for micronucleus induction by methyl methanesulfonate (MMS) and neocarzinostatin (NCS) both with and without the use of cytochalasin B in the micronucleus assay. Thus, defects in cellular responses to DNA damage are modulating factors in micronucleus formation. Topics: Animals; Ataxia Telangiectasia; Cells, Cultured; Cricetinae; Cricetulus; Cytochalasin B; DNA Damage; DNA Repair; Methyl Methanesulfonate; Micronucleus Tests; Mutagens; Nucleic Acid Synthesis Inhibitors; Radiation, Ionizing; Zinostatin | 1997 |
Dose-rate sparing for micronucleus induction in lymphocytes of controls and ataxia-telangiectasia heterozygotes exposed to 60Co gamma-irradiation in vitro.
We investigated the reproducibility of the cytochalasin B micronucleus (MN) assay in irradiated human lymphocytes to assess its suitability in predicting cancer predisposition and response to radiotherapy by virtue of defects in the processing of clastogenic lesions. G0 lymphocytes were exposed to 3.0 Gy 60Co gamma-rays at high (HDR) or low dose-rate (LDR). Six healthy donors were assayed three times each in nine experiments and compared with six ataxia-telangiectasia (A-T) heterozygotes. In controls, significant interexperiment variability in MN yields was observed at HDR and LDR, also in dose-rate sparing (i.e. reduction in MN yield at LDR compared with HDR). Significant inter-individual variability was seen at HDR, but not at LDR or for sparing. Average sparing was 66.4 +/- 4.8%. In spite of the experimental variability, a significant difference between controls and A-T heterozygotes was detected at LDR, and 5/6 heterozygotes had sparing values below the control range. This gives encouragement for the use of this assay in predictive testing if sources of experimental variability can be identified so as to improve discrimination between individuals. HDR and to a lesser extent LDR irradiation induced significant mitotic inhibition, seen as a reduction in binucleate cells after cytocholasin treatment. A positive correlation between mitotic inhibition and MN frequency suggests that similar lesions may be involved in these effects. Topics: Adult; Analysis of Variance; Ataxia Telangiectasia; Cobalt Radioisotopes; Cytochalasin B; Gamma Rays; Heterozygote; Humans; Lymphocytes; Male; Micronuclei, Chromosome-Defective; Micronucleus Tests; Middle Aged; Mitosis; Radiation Dosage; Reproducibility of Results | 1996 |
Response of fibroblast cultures from ataxia-telangiectasia patients to oxidative stress.
The object of this study was to determine whether ataxia-telangiectasia (AT) cells are more sensitive than normal cells to reduced oxygen species generated either during normal cell processes or resulting from metabolism of xenoblotics. To test this hypothesis four AT and four normal fibroblast cultures were exposed to hydrogen peroxide (H2O2) and the induction of micronucleated cells was assayed. AT cultures responded to the H2O2 treatment with a greater increase in micronucleus frequencies than that observed in normal cultures (P less than 0.01). At time course study showed that an elevation in micronucleus frequencies occurred earlier in AT cultures (significant increase by 1.5 h after treatment) than in normal cultures, possibly indicating a G2-phase sensitivity of AT cells to H2O2. The addition of an aqueous extract of areca nut to the cultures, as an example of exogenous stress, induced a greater frequency of micronucleated cells in AT cultures than in the normal cultures. These results suggest that the AT syndrome may serve as a model for investigating the role of reduced oxygen species in cancer. Topics: Areca; Ataxia Telangiectasia; Cell Division; Cell Line; Chromosome Aberrations; Cytochalasin B; Fibroblasts; Humans; Hydrogen Peroxide; Micronucleus Tests; Plant Extracts; Plants, Medicinal | 1990 |