FISH analysis of 1cen-1q12 breakage, chromosome 1 numerical abnormalities and centromeric content of micronuclei in buccal cells from thyroid cancer and hyperthyroidism patients treated with radioactive iodine.
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One of the health consequences of the Chernobyl nuclear power plant accident was a radioactive iodine-related increase in the incidence of thyroid cancer in exposed children. This radioisotope is used in the treatment of thyroid cancer and hyperthyroidism patients providing a convenient opportunity to study cytogenetic damage induced by known doses of radioactive iodine in treated patients. We used pancentromeric FISH on micronuclei and chromosome 1 tandem labelling FISH to monitor overall chromosome breakage and loss, 1q12 breakage and decondensation and chromosome 1 numerical abnormalities in buccal cells from 31 radioactive iodine-exposed hyperthyroidism and thyroid cancer patients. The overall outcome of the study, with 250,000 buccal cells analysed, is that there was no radioactive iodine-related increase in the frequency of micronuclei, 1q12 breakage, 1q12 decondensation or chromosome 1 numerical abnormalities. In addition, neither age nor gender, health status nor radioactive iodine dose modulated the frequency of the above cytogenetic end points. Although several uncertainties of these emerging molecular cytogenetic methodologies will require further experimentation, we conclude that, at the reported exposure levels, radioactive iodine did not induce detectable chromosome damage in buccal cells from treated patients. (+info)
Chromosome breakage and cell lethality in human hepatoma cells irradiated with X rays and carbon-ion beams.
(18/785)
Prediction of radiosensitivity would be valuable for heavy-ion radiotherapy. Premature chromosome condensation (PCC) technique has been a potential predictive assay in photon radiotherapy, but has not been investigated for hepatomas receiving heavy ions. Two human hepatoma cell lines, i.e., HLE and HLF, were irradiated with either 290 MeV/u carbon ions or 200 kVp X rays. Cell lethality was assayed by colony formation and compared with the unrejoined fraction of chromatin breaks as measured by PCC technique. Carbon ions at linear energy transfer (LET) of 76 keV/micron produced cell death more effectively than those of 13 keV/micron and X rays. For the cell killing, the relative biological effectiveness (RBE) of 13 and 76 keV/micron carbon ions compared with X rays was 1.10-1.24 and 2.57-2.59, respectively. Mean number of chromosomes in HLE and HLF cells was similar to each other, i.e., 60.48 and 60.28. RBEs for chromatin breaks of 13 and 76 keV/micron carbon ions were 1.30-1.31 and 2.64-2.79, respectively. A strong correlation between unrejoined chromatin breaks and cell killing for human hepatoma cells was observed irrespective of radiation quality. We conclude that PCC provides a potential predictor for the radiosensitivity of individual hepatoma that are treated with photon as well as heavy ion irradiation. (+info)
Chromosome breakage hotspots and delineation of the critical region for the 9p-deletion syndrome.
(19/785)
The clinical features of the 9p-deletion syndrome include dysmorphic facial features (trigonocephaly, midface hypoplasia, upward-slanting palpebral fissures, and a long philtrum) and mental retardation. The majority of these patients appear to have similar cytogenetic breakpoints in 9p22, but some cases show phenotypic heterogeneity. To define the breakpoints of the deleted chromosomes, we studied 24 patients with a deletion of 9p, by high-resolution cytogenetics, FISH with 19 YACs, and PCR using 25 different sequence-tagged sites. Of 10 different breakpoints identified, 9 were localized within an approximately 5-Mb region, in 9p22-p23, that encompasses the interval between D9S1869 (telomeric) and D9S162 (centromeric). Eight unrelated patients had a breakpoint (group 1) in the same interval, between D9S274 (948h1) and D9S285 (767f2), suggesting a chromosome-breakage hotspot. Among 12 patients, seven different breakpoints (groups 3-9) were localized to a 2-Mb genomic region between D9S1709 and D9S162, which identified a breakpoint-cluster region. The critical region for the 9p-deletion syndrome maps to a 4-6-Mb region in 9p22-p23. The results from this study have provided insight into both the heterogeneous nature of the breakage in this deletion syndrome and the resultant phenotype-karyotype correlations. (+info)
Absence of Brca2 causes genome instability by chromosome breakage and loss associated with centrosome amplification.
(20/785)
Women heterozygous for mutations in the breast-cancer susceptibility genes BRCA1 and BRCA2 have a highly elevated risk of developing breast cancer [1]. BRCA1 and BRCA2 encode large proteins with no sequence similarity to one another. Although involvement in DNA repair and transcription has been suggested, it is still not understood how loss of function of these genes leads to breast cancer [2]. Embryonic fibroblasts (MEFs) derived from mice homozygous for a hypomorphic mutation (Brca2(Tr2014)) within the 3' region of exon 11 in Brca2 [3], or a similar mutation (Brca2(Tr)) [4], proliferate poorly in culture and overexpress the tumour suppressor p53 and the cyclin-dependent kinase inhibitor p21(Waf1/Cip1). These MEFs have intact p53-dependent DNA damage G(1)-S [3] [4] and G(2)-M checkpoints [4], but are impaired in DNA double-strand break repair [3] and develop chromosome aberrations [4]. Here, we report that Brca2(Tr2014/Tr2014) MEFs frequently develop micronuclei. These abnormal DNA-containing bodies were formed through both loss of acentric chromosome fragments and by chromosome missegregation, which resulted in aneuploidy. Absence of Brca2 also led to centrosome amplification, which we found associated with the formation of micronuclei. These data suggest a potential mechanism whereby loss of BRCA2 may, within subclones, drive the loss of cell-cycle regulation genes, enabling proliferation and tumourigenesis. (+info)
Excision of micronuclear-specific DNA requires parental expression of pdd2p and occurs independently from DNA replication in Tetrahymena thermophila.
(21/785)
Elimination of germ-line DNA segments is an essential step in the somatic development of many organisms and in the terminal differentiation of several specialized cell types. In binuclear ciliates, including Tetrahymena thermophila, DNA elimination occurs during the conversion of the germ-line micronuclear genome into the somatic genome of the new macronucleus. Little is known about molecular determinants and regulatory mechanisms involved in this process. Pdd2p is one of a small set of Tetrahymena polypeptides whose time of synthesis, nuclear localization, and physical association with sequences destined for elimination suggest an involvement in the DNA elimination process. In this study, we report that loss of parental expression of Pdd2p leads to the perturbation of several DNA rearrangement processes in developing zygotic macronuclei, including excision of internal eliminated sequences, excision of chromosome breakage sequences, and endoreplication of the new macronuclear genome and eventually results in lethality of the progeny. We demonstrate that excision and elimination of micronuclear-specific DNA occurs independently of endoreplication of the new macronuclear genome that takes place during the same period of time. Thus, our data indicate that parental expression of Pdd2p is required for successful DNA elimination and development of somatic nuclei. (+info)
Evaluation of micronuclei and chromosomal breakage in the 1cen-q12 region by the butadiene metabolites epoxybutene and diepoxybutane in cultured human lymphocytes.
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1,3-Butadiene is a widely used industrial chemical and common environmental pollutant that has been associated with increased risks of leukemias and lymphomas. Butadiene and its metabolites, 1, 2-epoxybutene (EB) and diepoxybutane (DEB), have been shown to be genotoxic in a wide variety of test organisms. The objective of this research was to evaluate techniques for the rapid detection of chromosomal alterations occurring in humans exposed to butadiene. We have used a multicolored fluorescence in situ hybridization (FISH) method and the CREST-modified micronucleus assay to detect chromosomal breakage induced by EB (10-300 microM) and DEB (0.5-10 microM) in cultured human lymphocytes. A significant dose-related increase in the formation of micronuclei was seen in lymphocytes treated with DEB at concentrations as low as 2.5 microM, but not with EB over the dose range tested. Over 80% of the micronuclei induced by DEB were CREST-negative, indicating their origin from chromosomal breakage. Multicolor FISH using two adjacent chromosome-specific probes showed a significant increase in chromosomal breakage in the 1cen-q12 region induced by DEB at concentrations as low as 2.5 microM, but not by EB. Since DEB is likely to be one of the metabolites contributing to the genotoxic effects of butadiene, the sensitivity of the tandem FISH approach to detect breakage induced by diepoxybutane indicates that this technique may be useful for monitoring chromosomal alterations in butadiene-exposed workers. (+info)
A 28-kb deletion spanning D15S63 (PW71) in five families: a rare neutral variant?
(23/785)
Methylation analysis with probe PW71 (D15S63) is an established procedure to test patients suspected of having Prader-Willi syndrome or Angelman syndrome. Using this test, we have identified a 28-kb deletion spanning D15S63 in five independent families. Sequence analysis revealed identical breakpoints in all the families. The haplotype data are compatible with a common ancestral origin of the deletion in at least two families. The deletion was not found in 1, 000 unrelated controls. Although the deletion maps within the imprinting-center region, neither maternal nor paternal inheritance of the deletion appears to affect imprinting in proximal 15q. We conclude that the deletion is a rare neutral variant that can lead to false-positive results in the PW71-methylation test. (+info)
Clustered 11q23 and 22q11 breakpoints and 3:1 meiotic malsegregation in multiple unrelated t(11;22) families.
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The t(11;22) is the only known recurrent, non-Robertsonian constitutional translocation. We have analyzed t(11;22) balanced-translocation carriers from multiple unrelated families by FISH, to localize the t(11;22) breakpoints on both chromosome 11 and chromosome 22. In 23 unrelated balanced-translocation carriers, the breakpoint was localized within a 400-kb interval between D22S788 (N41) and ZNF74, on 22q11. Also, 13 of these 23 carriers were tested with probes from chromosome 11, and, in each, the breakpoint was localized between D11S1340 and APOA1, on 11q23, to a region +info)