Chromosomal breakage and sister chromatid exchange in peripheral blood lymphocytes and lymphoblastoid cell lines in the X-linked lymphoproliferative syndrome

The frequencies of chromosomal aberrations and sister chromatid exchange (SCE) were measured in lymphoblastoid cell lines (LCLs) and in phytohemagglutinin (PHA)- and pokeweed mitogen (PWM)-stimulated lymphocytes from males with X-linked lymphoproliferative (XLP) syndrome, their obligate carrier mothers, and control subjects. We observed an increased frequency of chromosomal aberrations including increased polyploidy in LCLs derived from families with XLP with time in culture. The SCE rate in LCLs (mean of 3.89 SCEs per cell) was much lower than that in PHA- or PWM-stimulated lymphocytes: PWM-stimulated lymphocytes showed 9.58 SCEs per cell and PHA-stimulated cells had 11.38 SCEs per cell. A greater number of chromosomal gaps and breaks in the D-group chromosomes of LCLs of affected males and carrier females were identified compared to the number expected, based on chromosomal length and the number of aberrations seen in PHA-stimulated cell cultures. No differences in the frequency of SCEs or chromosomal aberrations were found in control subjects and affected males or carrier females in the peripheral lymphocytes stimulated by PHA. Phenotypes of XLP appear to arise from failure of immune responses to Epstein-Barr virus (EBV) and not from intrinsic chromosomal breakage or instability.     

Elevated sister chromatid exchange and cell cycle analysis in bone marrow in childhood ALL

Sister chromatid exchange (SCE) frequencies were analyzed in both stimulated and unstimulated bone marrow samples from eight patients recently found to have childhood acute lymphocytic leukemia (ALL). Six of the patients had elevated SCE frequencies in stimulated marrow when compared to control values. In unstimulated marrow, all four patients had elevated SCE frequencies. These data agree with peripheral leukocyte findings in ALL. Cell cycle analysis revealed no significant differences between the patient marrows and the control marrows. Since no correlation between cell cycle distribution and SCE frequency was found, it is suggested that the differences in SCE observed may be related to the type of proliferating cell.     

Elevated sister chromatid exchange rate in lymphocytes of the European eel (Anguilla anguilla) with cauliflower tumor

Sister chromatid exchange (SCE) frequency and chromosomal aberration were studied in cultured lymphocytes from the European eel (Anguilla anguilla) with cauliflower tumors. SCE was significantly increased in the affected fishes compared with normal controls, whereas chromosomal aberration did not. In comparison to normal cells, the lymphocytes responding to phytohemagglutinin M (PHA-M) in cultures from tumorous eels showed no evidence of cell cycle retardation. A possible mechanism for elevation of SCE and the significance of this finding relation to the etiology were discussed.     

Some comments on sister chromatid exchange (SCE) in human neoplasia

The incidence of sister chromatid exchange (SCE) in bone marrow cells and/or lymphocytes of patients with various leukemias and the effects of drugs on the SCE incidence in the cells of patients with leukemia or cancer are presented and discussed. The possible use of SCE for screening antileukemic drugs, mutagenic and/or carcinogenic agents and susceptible human populations is presented.     

Evidence that the replication point is the site of sister chromatid exchange

DNA synthesis in Chinese hamster cells was blocked partially by treating the cells with either fluorodeoxyuridine (FUdR) or cycloheximide (CHM) for various lengths of time. Analyses of the population kinetics and measurement of incorporation of labeled nucleosides during the FUdR block strongly suggested that the number of growing points was accumulated by the treatment while the rate of chain growth was greatly reduced. No evidence for such an accumulation was obtained in the CHM-treated cells. To study the relation between DNA replication and sister chromatid exchange (SCE), bromodeoxyuridine-labeled cells were exposed to blue fluorescent light while DNA synthesis was blocked. The frequency of SCE induced by the light treatment appeared to increase as the number of growing points increased, implying that the site of exchange is confined to the replication forks. The induction of SCE by fluorescent light was inhibited completely by CHM-treatment. The reason for this finding remains to be elucidated.     

Temperature-dependence of sister chromatid exchange: An implication for its mechanism

To obtain some insight into the mechanism for spontaneous sister chromatid exchange (SCE), the effect of temperature on the incidence of SCE was studied by culturing Chinese hamster cells at various temperatures. The frequency of SCE increases fivefold when the incubation temperature was raised from 31 to 42°C; this increase was strictly temperature dependent. An Arrhenius plot of the SCE frequency indicated that the formation of SCE was enhanced greatly when the temperature was raised above 39°C. Cell growth at temperatures higher than 39°C also caused a marked elongation of the DNA synthetic phase. A working hypothesis was proposed that spontaneous SCE might result from a cooperative of processes involved in repair of spontaneous DNA damage and DNA replication.     

Sister chromatid exchange levels and cell cycle time in human bone marrow cells and lymphocytes

The frequency of sister chromatid exchange (SCE) and the cell-cycle-specific pattern of mitoses were analyzed at the same time in normal bone marrow cells and lymphocytes of six healthy donors. The SCE frequency was found to be significantly higher in lymphocytes. The cell-cycle-specific pattern revealed significantly shorter cell cycle times for normal bone marrow cells as compared with those of phytohemagglutinin (PHA) stimulated lymphocytes. Chromosomes of bone marrow metaphases displayed a more contracted morphology.     

Chromosome damage and sister chromatid exchanges in lymphocyte cultures from patients with two primary cancers

Sister chromatid exchanges (SCEs) and chromosome damage were scored in lymphocyte cultures from 11 patients with two or more primary cancers and were compared with normal controls. None of the patients had a constitutional chromosome anomaly, but six showed evidence of chromosome instability, which could not be accounted for by treatment, expressed either as elevated SCE frequency or increased nonspecific chromosome damage and chromosome loss. Chromosome damage included major rearrangements as well as deletions and gaps. The possibility of common mechanisms in chromosome instability leading to susceptibility to a heterogeneous group of primary cancers is discussed.     

Chromosome replication in normal and transformed human lymphocytes

We analyzed the late-replication patterns of human B-lymphocyte chromosomes before and after transformation by Epstein-Barr virus. There were no statistically significant differences between normal cells and transformed cells derived from the same male individual; therefore, the order of termination of chromosome replication was unchanged by transformation. We also examined the replication patterns of T lymphocytes from the same donor and found no differences between normal B and T cells.     

Malignant melanoma: sister chromatid exchange analysis in three families

Sister chromatid exchange (SCE) was analyzed in stimulated lymphocytes and skin fibroblasts in members of three families with cutaneous malignant melanoma (CMM). Two of these families were characterized by familial CMM; the other family had one patient affected by CMM and two others with other cutaneous melanocytic lesions. All the patients had undergone surgery but no chemotherapy. Higher and differing SCE rates were found in lymphocytes and in fibroblasts of all patients. A wide range of SCE distribution was found in patients with high SCE rate. A few healthy close relatives also showed relatively high SCE rates and wide range distributions. These subjects may be regarded as a subset of family members at high risk for developing cancer. The variability of SCE rates and distribution may reflect genetic heterogeneity of CMM.     

Evidence for an indirect effect of radiation on mammalian chromosomes. III. UV- and x-ray-induced sister chromatid exchanges in heterokaryons

The hypothesis that chromosomes may be damaged indirectly by radiation was examined by assaying sister chromatid exchange, (SCE) frequency in heterokaryons between irradiated and unirradiated mouse and Chinese hamster cells. One cell line was UV or x irradiated, then fused to unirradiated BrdU-labeled cells of the other line; SCEs in the unirradiated set were scored in heterokaryons. A dose-dependent increase was consistently observed; the magnitude of which suggested that 25% of UV-induced and up to 60% of x-ray-induced SCEs are indirectly induced. Medium transfer experiments, cell mixing, and fusion with irradiated chick erythrocyte ghosts suggested that unirradiated chromosomes in heterokaryons are damaged by a stable, nondiffusible cytoplasmic component contributed by the irradiated cell.     

Sister chromatid exchange: Variation by age,sex, smoking,and breast cancer status

Variation in sister chromatid exchange (SCE) frequency in lymphocytes of 125 persons was compared using a multivariate general linear model. The study was performed to determine whether SCE frequency differs with respect to age, sex, smoking, and breast cancer status. Study subjects were divided into: members of two branches of families having an excess of cancer (primarily breast) including a brother and sister in one family who developed nonbreast malignancies within 1 yr of the study; women in both families successfully treated for breast cancer (all at least 5 yr posttreatment); and women from the general population with confirmed breast cancer.Controls consisted of spouses who married into the high-risk kindreds, hospital personnel, and others (primarily tradesmen without history of occupational exposure). Results show that: (1) Women with active breast cancer have a significantly higher mean SCE frequency than control women or women greater than 5 yr posttreatment for breast cancer; (2) Cigarette smokers show a significantly higher number of SCEs than was observed in nonsmokers; (3) The increase in SCE level in smokers is dose-related to exposure as measured by cumulative pack-years; (4) SCE values in both high-risk families are not significantly different from controls; (5) Neither the age nor sex of the individual affects SCE frequency; and (6) The observed distribution of exchanges agrees with that expected based on the proportion of the genome represented by each chromosome group.     

Sister chromatid exchange in aplastic anemia

The incidence of sister chromatid exchanges (SCE) in the lymphocytes of patients with aplastic anemia (AA) was determined before and after exposure to mitomycin C (MMC). The “baseline” SCE rate was significantly higher in AA, but MMC-induced SCE rate was not different compared to controls. It is suggested that some patients with AA may have an underlying DNA damage.     

Sister chromatid exchange in normal and Ph1-positive leukemic cells after mitomycin-C treatment in vitro

The sister chromatid exchange (SCE) frequency was investigated in normal bone marrow and Ph1-positive cells of chronic myelocytic leukemia (CML) patients with and without mitomycin-C (MMC) treatment in vitro. Even though the spontaneous SCE frequency was found to be significantly lower in CML cells, the absolute SCE values after MMC treatment did not differ between leukemic and normal cells, and this seems to indicate an equilization of SCE rates. However, the fact that leukemic cells with lower spontaneous SCE rates need a further increase of SCE to reach values equal to those of normal cells might indicate a somewhat higher susceptibility of leukemic cells to DNA damage by MMC. This interpretation appears to be confirmed by the fact that the inhibition of cellular proliferation at higher MMC doses considerably reduced the number of leukemic cells that was able to divide twice during a given culture time.     

Baseline and mitomycin-c-induced sister chromatid exchanges in a melanoma and a colon tumor cell line

Baseline and mitomycin C (MMC)-induced sister chromatid exchanges (SCEs) in two human tumor cell lines (a colon tumor and a melanoma) and in a normal fibroblast cell line were analyzed and compared. The tumor cells showed numerical and structural chromosomal abnormalities. Their baseline SCE rate was slightly, but not significantly, higher than that of the control. Each tumor cell line showed a dose-dependent increase in SCE frequency above the spontaneous level in its own specific manner. The response in the malanoma cell was consistantly below that of the control, but only the response to the highest dose of MMC (10^?9 M) was significantly lower than that of the control. The response of the colon tumor cells varied with respect to that of the control. Thus, it appears that karyotypic instability in tumor cells is not necessarily associated with elevated baseline or induced SCE/chromosome rates. In addition, within each cell line dose group, the SCE frequency was proportional to the number of chromosomes. Thus, the SCE/chromosome is a better expression of genetic damage than SCE/metaphase in analyses involving heteroploid cells.     

Mapping of amplified c-myb oncogene, sister chromatid exchanges, and karyotypic analysis of the COLO 205 colon carcinoma cell line

We have studied molecular and chromosomal details of cytogenetic status in a human tumor cell line COLO 205 that shows a stable, approximately tenfold amplification of the c-myb oncogene. The amplified copies of c-myb reside in two marker chromosomes that may have evolved from chromosome #6 by complex chromosomal rearrangements. No homogeneously staining regions can be discerned at the site of c-myb amplification. We suggest that c-myb was amplified in situ in a chromosomal segment (6q22-24) that became a part of the marker chromosome, possibly through isochromosome formation followed by duplication, and without the extrachromosomal intermediate form of double minute chromosomes. There is an enhanced frequency of sister chromatid exchanges at the site of amplified c-myb. These results are discussed in the context of models for gene amplification and oncogene activation.     

High frequency of in vivo sister chromatid exchanges in the bone marrow cells of mice bearing mammary adenocarcinoma dbrB

Sister chromatid exchanges (SCEs) in vivo were evaluated in the bone marrow cells of mice bearing mammary adenocarcinoma dbrB of 0.5 and 1.0 cm³, respectively. It has been observed that the SCE frequency is significantly higher (p < 0.001) in the bone marrow cells of tumor-bearing mice as compared to the bone marrow cells of non-tumor-bearing mice. SCE frequency in the bone marrow cells of mice with 1.0-cm³ tumors did not increase appreciably with tumor size as compared to that in the bone marrow cells of mice with o.5-cm³ tumors. This indicates the presence of mammary carcinoma-induced somatic stress as noted in the genetic material of the bone marrow cells of the host.     

The structure and origin of giant nuclei in human cancer cells

The structure and origin of giant nuclei in human cancer cells were studied in two cases of squamous cell carcinoma of the cervix. Feulgen-squash preparations of untreated biopsies showed the following types of nuclei: fairly evenly stained nuclei, nuclei with distinct chromocenters from which individual chromosomes sometimes seemed to radiate, and nuclei with banded structures which we tentatively interpret as some type of “polytene” chromosomes possibly showing prophase chromomeres. In neither tumor did either the small or large nuclei display any X chromatin bodies. The main processes giving rise to the different types of nuclei are endoreduplication, true endomitosis, and a kind of polytenization. The multichromosome chromocenters that fell apart possibly represent polytene chromosomes separating into individual chromosomes.     

Indirect stimulation of B-cell proliferation in vitro by T cells,as evidenced by cytogenetic analysis of PHA-stimulated cell cultures of B-cell lymphomas

In a retrospective study of non-Hodgkin's lymphomas, the 14q+ marker was found in at least one of the samples examined from 17 patients with B-cell lymphoproliferative diseases (LPD). In the PHA-stimulated cultures, the marker was found in each sample in 10%–100% of the cells. An indirect stimulation, as indicated by a ³H-thymidine incorporation and IG secretion, of normal B cells by a T-cell mitogen, such as PHA, has been recently documented. This phenomenon is confirmed by our chromosome analysis, which demonstrated characteristic chromosome changes in PHA-stimulated cultures of patients with B-cell malignancies and indicated that the phenomenon can be observed not only in normal B cells but also in malignant B cells.