Appearance of altered cell-surface fucosyl glycopeptides in concomitance with chromosomal alterations in the gross virus-infected pre-leukemic thymus of the rat

The appearance of a class of fast-eluting cell-surface glycopeptides that are encountered almost exclusively in malignant and certain pre-malignant cells was monitored in the course of leukemogenesis in the thymus of rats injected at birth with Gross leukemia virus. The altered glycopeptides appeared as early as 15 days after virus injection, when the animals were still clinically healthy and no histological signs of the disease were present in the thymus. Their amount was further increased at 30 days, and reached a maximum in the fully developed lymphoma. The development of this early phenotypic marker of malignancy appeared to be concomitant with that of chromosomal anomalies in the thymus. Since these anomalies are non-random, the existence of a causal relationship between the glycopeptide change and the loss of specific chromosomes might be hypothesized

Chromosome fragile sites in Down syndrome patients

We report on the fragile site expression in lymphocytes from 16 Down syndrome (DS) men aged 24 to 52 years. Among rare fragile sites, 16q22 has been reported to be induced or enhanced by alpha-interferon. Since DS cells have three copies of the receptor for alpha-interferon, we hypothesized a possible enhancement of 16q22 expression. This fragile site has also been related to a specific rearrangement in M4 acute nonlymphocytic leukemia. In view of the high incidence of acute leukemia in DS subjects, we studied the expression of 16q22 in lymphocyte cultures treated with 5-bromodeoxyuridine or alpha-interferon. We also studied whether the repair deficiency of DS cells could affect the expression of aphidicolin-induced fragile sites. The level of chromosomal aberrations was compared with that found in aphidicolin-treated cultures from 12 normal subjects of the same age. We found neither spontaneous or BrdU-induced fragility at 16q22 nor induction by alpha-interferon. Chromosomal breakage rate was increased in alpha-interferon-treated cultures in comparison with control cultures of the same subjects. Aphidicolin-induced fragile sites expression in DS patients did not differ significantly from that found in the lymphocyte cultures from control subjects

Do human lymphocytes exposed to the fallout of the Chernobyl accident exhibit an adaptive response? III. Challenge with bleomycin in lymphocytes from children hit by the initial acute dose of ionizing radiation

In the present paper, we report data on the possible adaptive response, induced in vivo by exposure to ionizing radiation to a challenge treatment with the radiomimetic glycopeptide bleomycin (BLM). Lymphocytes from children living in Pripjat at the time of the Chernobyl accident, and thus hit by the initial acute dose of ionizing radiation, were treated for the last 5 h of culture with 0.004 U/ml BLM. Significantly lower chromosome damage was found only in lymphocytes from children who, independently of the initial acute exposure to ionizing radiation, still showed a 137Cs internal contamination, due to persistent continuous exposure to low doses of radiation. The present results indicate that past exposure to acute high dose of ionizing radiation does not interfere with resistance to BLM which is related to internal contamination

Association of cytogenetic abnormalities in a neuroblastoma and fragile sites expression

A 15 month old boy with a stage IV right suprarenal gland neuroblastoma showed a number of raised biochemical parameters, whilst catecholamines and skeletal survey were normal. Treatment with peptichemio failed to give a clinical response. Histological evidence of neuroblastoma infiltration in the bone marrow aspirate was absent. Immunofluorescence on sedimented cells was negative using antibody UJ223.8, PI153/3 and H11; only UJ308 and to a lesser extent UJ13A gave positive results. After 21 days, however, the same cells in culture showed highly differentiated dendritic processes. Thirty-seven percent metaphases from bone marrow aspirate showed the following karyotype 45XY, del (1) (p32), and two markers. Mar1 = der (2) t (2; 2) (2qter→2q14::2p24→2qter). Mar2 = der (15) t (15; 2) (15qter→15p11::2p11→2pter). Treatment with methotrexate reduced the aberrant mitoses rate to 2%. N-myc in situ hybridisation showed significant signal on both markers confirming the cytogenetic interpretation. Peripheral blood lymphocytes at 72 h showed a higher level of breaks per cell than control. After treatment with aphidicolin (APC) or methotrexate (MTX) for the last 24 h, to induce fragile sites, the incidence of breaks per cells was increased. Moreover 11.4% of APC-induced breaks were in 1p31-32 (mean of normal controls = 2.3%). The mother presented an increased sensitivity to the inducibility of fragile sites, while the father's lymphocytes showed values within the control range. The genetic changes produced by the abnormalities on chromosomes 1 and 2 might be related to tumour progression. Furthermore th is is the first description of correlation between a high frequency of fragile site 1p31-32 induced by APC in the patient's lymphocytes and deletion of 1p32 in tumour cells. The interpretation of these findings and of other similar correlations needs further study

DNA damaging agents can induce celll cycle arrest in different phases of mitosis

DNA damaging drugs often induce cell cycle arrest at G1 or G2, to allow the repair of damage, or apoptosis induction. Mitotic checkpoints have been also associated to DNA damage response, even if they are less well characterized. In this study we analyzed the progression through mitosis of the mismatch repair deficient (MMR-) HCT116 and HCT15 colon cancer cell lines and their MMR proficient (MMR+) counterparts, after treatment with the DNA polymerase inhibitor aphidicolin, the radiomimetic bleomycin and the Topoisomerase I (Topo) I inhibitor SN38. Cells were treated with the different drugs at concentrations and times suitable to detect DNA and chromosome damage, without abrogating mitosis. After fixation, cells were incubated with anti-α, -β or -γ tubulin antibodies and a secondary fluorescent antibody; DNA was counterstained with DAPI. The percentage of cells in the various phases of mitosis, abnormal mitoses (multipolar, with lagging chromosomes, etc.) and mitotic indexes were evaluated. Aphidicolin and bleomycin caused cell arrest mainly at the checkpoint between metaphase and anaphase, whereas SN38 induced cell accumulation at the end of mitosis, when tubulins are present only in the midbodies, just before the definitive cytodieresis. Since Aurora kinase B is essential in the control of cytodieresis, we plan to analyze the possible intervention of this kinase in tumor cell response to the drugs under study. Moreover, we are evaluating telomere damage that can be induced by Topo I inhibitors and may cause cell arrest at the end of mitosi

Aphidicolin and bleomycin induced chromosome damage as biomarker of mutagen sensitivity: a twin study

The induction of chromosome damage in cultured human lymphocytes by in vitro treatments with aphidicolin (APC) and bleomycin (BLM) has been proposed as test of sensitivity to mutagens. To assess their validity, we have investigated whether the individual expression of induced chromosome damage has a genetic rather than an environmental basis. Metaphase analysis for chromosomal aberrations (CA) and micronucleus (MN) assay in cytokinesis-blocked cells have been performed in peripheral blood lymphocytes from 19 healthy male twins (9 monozygotic and 10 dizygotic pairs), aged 70-78 years, after APC, BLM and APC+BLM treatments. Concordance between twins revealed a high genetic component in the sensitivity towards clastogenic action of APC both as percentages of CA and MN. The micronucleus assay demonstrated a genetic basis also in the expression of chromosome damage induced by BLM and APC+BLM treatments. Since twins were elderly people, to investigate the possible role of age, CA and MN frequencies were compared with those found in lymphocytes from 11 young male donors. Basal and APC-induced chromosome damage were clearly increased in the former. Following BLM and APC+BLM treatments, age significantly increased mitotic delay, as shown by the mitotic indexes (MI) and by the ratios between binucleated and mononucleated (B/M) cells. © 2003 Elsevier B.V. All rights reserved

Cytogenetic effects of 1-p-(3-methyltriazeno)benzoic acid potassium salt on human lymphocytes in vitro

The triazene derivative 1-p-(3-methyltriazeno)benzoic acid potassium salt (MTBA) shows pharmacological properties similar to those of 5-(3,3-dimethyl-1-triazeno)imidazole-4-carboxamide (DTIC, trade name dacarbazine), which is known to induce antigenic modulation in tumor cells (xenogenization) and is currently used in cancer therapy. Mutagenic, teratogenic and cancerogenic properties of triazene derivatives have been demonstrated but there is no report on their possible clastogenicity. We describe here the in vitro cytogenetic effects of MTBA on human peripheral blood lymphocytes. The drug was tested at different culture times in a range of concentrations from 2 to 500 micrograms/ml. MTBA caused a dose-dependent increase in the frequency of chromosomal breaks. Different blood donors showed different sensitivity to the treatment. Cell proliferation, as evaluated by [3H]thymidine incorporation, was inhibited at the highest concentrations of the drug. These data might be relevant for comparison with in vivo effects of the drug in clinical trials and to investigate the possible relations between xenogenization induced by MTBA and its genetic and cytogenetic effects in human lymphocytes