Sister Chromatid Exchanges In Human Lymphocytes Research Articles

In vitro genotoxic effects of the anticancer drug gemcitabine in human lymphocytes

This research was carried out to investigate in vitro genotoxic effects of the anticancer agent gemcitabine on the induction of chromosomal aberrations and sister-chromatid exchange in human lymphocytes. Three doses of gemcitabine (0.001, 0.002 and 0.004 μg/ml) were applied to lymphocyte cultures from 15 donors. There was a significant increase in the induction of chromosome aberrations and in the occurrence of sister-chromatid exchange in these cells. In addition, gemcitabine significantly decreased the mitotic index and replicative index for all doses. Dose–response regression lines were used to compare the individual susceptibilities to gemcitabine with respect to the chromosome aberration and sister-chromatid exchange frequencies. Our results indicate that gemcitabine is able to induce both cytotoxic and genotoxic effects in human lymphocyte cultures in vitro in a dose-dependent manner.

Effects of donor age and contamination level of dioxins and related chemicals on frequency of sister chromatid exchanges in human lymphocytes cultured in vitro

Frequencies of sister chromatid exchanges (SCEs) of human lymphocytes in control (DMSO treated) and 7,8-benzoflavone (ANF) treated cultures were measured in 39 healthy Japanese people and examined in connection with donor age. Both the control (baseline) and ANF induced SCE rates were significantly enhanced with age and highly positive correlation was observed between them. Therefore, in vivo aging seemed to have some effects on the frequencies of SCEs in human lymphocytes cultured in vitro. Concentrations of PCDDs, PCDFs and Co-PCBs in the blood and sebum of face were determined in the same Japanese subjects. Significantly positive correlation was observed between the blood and sebum in their total TEQ levels. Hence, PCDDs, PCDFs and Co-PCBs, which have been contaminating human bodies, are considered proportionally excreted from the sebum of face or body. Their total TEQ concentrations were also meaningfully increased with donor age in the sebum of face as well as in the blood. Either the baseline or ANF induced SCE frequencies was not enhanced with the total TEQ levels in the blood. Therefore, background levels of their contamination seem not to affect the SCE rates of the lymphocytes in the control and ANF treated cultures.

Amino acids excess increase SCEs in human lymphocytes

The effects of 21 amino acids on sister-chromatid exchanges (SCEs) in human lymphocytes were investigated. Addition of each of the 21 amino acids, originally in a composition of the culture medium RPMI 1640, at three different concentrations (10 μg/ml, 50 μg/ml and 100 μg/ml) caused a dose-independent increase in SCE frequency. The results indicated that amino acids, although indispensable for cells in vivo and in vitro, may be SCE-inducing agents.

Cytogenetic effects of a food mutagen, 2-amino-1-methyl-6-phenylimidazo[4,5- b]pyridine (PhIP), and its metabolite, 2-hydroxyamino-1-methyl-6-phenylimidazo[4,5- b]pyridine ( N-OH-PhIP), on human and Chinese hamster cells in vitro

2-Amino-1-methyl-6-phenylimidazo[4,5- b]pyridine (PhIP) induced structural chromosomal aberrations (CAs) and sister-chromatid exchanges (SCEs) in human lymphocytes and human diploid fibroblasts (TIG-7) at concentrations above 12.5 μg/ml in the presence of rat S9 mix. PhIP also elevated the frequencies of SCEs in human lymphocytes in the presence of rat S9 at concentrations above 2.0 μg/ml with dose-dependency. A proximate form of metabolites of PhIP, 2-hydroxyamino-1-methyl -phenylimidazo[4,5- b]pyridine ( N-OH-PhIP), caused CAs in human and Chinese hamster fi fibroblast cells in the absence of S9 mix at concentrations above 0.75 μg/ml and 1.25 μg/ml, respectively, which were 10 times lower than the effective concentration of PhIP. No marked differenceswere observed in the cytogenetic sensitivity to N-OH-PhIP between human and Chinese hamster cells, except between lymphocytes obtained from different donors.

Sister chromatid exchange in human lymphocytes induced by propoxur following plant activation by Vicia faba.

Because the carbamate insecticide propoxur induced sister chromatid exchanges (SCE) in Vicia faba but was ineffective in producing SCE in lymphocytes in culture, it was hardly suspected that plant metabolism was involved. Experiments were conducted in which metabolic activation was afforded by Vicia faba roots, and SCE in human lymphocytes in vitro was used to assess cytogenetic damage. Several concentrations of propoxur (250, 500, 1,000, 1,500, and 2,000 ppm) were applied for 4 hr to the roots of Vicia faba. Extracts prepared from these treatments were added to the lymphocyte cultures and a significant increase of SCE frequencies with a concentration-response relationship could be detected. The lymphocyte proliferation kinetics and the proliferation rate index (PRI) were not affected (except in the highest concentration, of 2,000 ppm). This general behavior was in agreement with the presence of an enzymatic system (S10 fraction) in Vicia roots capable of metabolizing or activating the propoxur. With 2,000 ppm, cell necrosis was produced in Vicia; therefore, this extract did not induce SCE in lymphocytes. However, lymphocyte proliferation kinetics were delayed and PRI was significantly decreased. Ethanol, a promutagen activated by this plant, was applied directly to the lymphocyte cultures as a positive control, and the response was negative. On the other hand, the extracts of roots treated with ethanol increased the SCE to more than twice that of the negative control, but the lymphocyte proliferation kinetics and PRI were not affected.

Effect of oxygen concentration in culture on the frequency of sister-chromatid exchanges in human lymphocytes and CHL cells

Effect of oxygen concentration in culture on the frequency of sister-chromatid exchanges in human lymphocytes and CHL cells

Sister-chromatid exchanges in human lymphocytes induced by dimethoate, omethoate, deltamethrin, benomyl and their mixture

Dimethoate and omethoate, two common organophosphorus insecticides, induced a dose-related increase inthe frequency of siter-chromatid exchanges (SCEs) in human lymphocytes in vitro ( P of the regression lines < 0.01). Two other common pesticides, the pyrethroid insecticide deltamethrin and the systemic fungicide benomyl, induced a modest increase in SCEs which bordered on statistical significance ( P = 0.053 and 0.055, respectively). Mixtures of the four pesticides at total concentrations of 41.5 and 83 μg/ml (composed of 43% dimethoate, 43% omethoate, 12% deltamethrin and 1.2% benomyl) induced a dose-dependent increase in SCEs ( P < 0.01). The effects of these mixtures of pesticides were variable using lymphocytes from different individuals, although these difference did not attain statistical significance. Moreover, low concentrations of the four pesticides that did not increase SCEs significantly when tested alone, were positive for SCE induction when tested as a mixture. The experiments show that sub-threshold doses of pesticides may increase SCEs when present in a mixture.

Genotoxicity of tauromustine, a new water soluble taurine-based nitrosourea. I. Mutagenic and clastogenic activity of tauromustine in vitro

Tauromustine (TCNU) a new taurine-based nitrosourea in phase III clinical trials against colon cancer, has been tested and compared with 1-(2-chloroethyl)-3-(4-methylcyclohexyl)-1-nitrosourea (CCNU) for mutagenic potential in Salmonella typhimurium TA98 and TA100 and, for its ability to cause chromosome aberrations and sister chromatid exchanges (SCE) in human lymphocytes. A dose dependent increase in the number of mutations in S. typhimurium TA100 but not in TA98 was found from 1.6 to 1000 micrograms/plate for both TCNU and CCNU. In addition, in the presence of Aroclor activated liver microsomes, CCNU caused a further increase in the mutation frequency which was not found for TCNU. A dose dependent increase in the chromosome aberration rate in human lymphocytes was induced by both TCNU and CCNU at concentrations ranging from 3.75 to 30 micrograms/ml after 24 h treatment during the last cell cycle. When lymphocytes were treated in G0 both TCNU and CCNU induced a higher frequency of aberrations at 15 and 30 micrograms/ml than during the last cell cycle and, unexpectedly, chromosome-type, i.e. dicentrics appeared. The difference in the frequency of aberrations in these two phases may be related to the different levels of O6-methyl guanine transferase present in resting contra cycling lymphocytes. TCNU at 0.8-8 micrograms/ml also induced a dose dependent increase in the number of SCE in human lymphocytes. Neither the number of chromosomal aberrations nor the number of SCE were affected by the addition of 1000 micrograms/ml taurine to the culture medium.

Genotoxicity testing of extracts of a Swedish moist oral snuff

The present study was designed to investigate the potential genotoxicity of aqueous and methylene chloride extracts of Swedish moist oral snuff. The test systems were selected to provide optimal data for the prediction of carcinogenicity in rodents and included assays for the induction of mutation in bacteria, sister-chromatid exchanges (SCE) in human lymphocytes, of chromosome aberrations and gene mutations in V79 Chinese hamster cells and of micronuclei in mouse bone marrow cells. In addition, the methylene chloride extract was tested for the induction of sex-linked recessive lethal mutations in Drosophila melanogaster. The aqueous extract of ‘Snus’ induced SCE in human lymphocytes and chromosome aberrations in V79 cells, the latter effect being observed both with and without metabolic activation. No induction of point mutations was detected with the Ames test or in V79 cells and the micronucleus test in mice was negative. It was demonstrated that the induction of chromosome aberrations without metabolic activation may be due to a high salt concentration, indicating that the clastogenic agent(s) in this extract required metabolic activation. The methylene chloride extract showed genotoxicity in the Ames test, the SCE test and the chromosome aberration test, whereas no induction of gene mutations in V79 cells was observed. Once again, the results suggested that metabolism is required for genotoxicity. The methylene chloride extract did not cause induction of micronuclei in mice or of sex-linked recessive lethal mutations in Drosophila melanogaster. These combined data on genotoxicity were analyzed using various models for the prediction of carcinogenicity. In a sequential testing model, the probabilities that the aqueous and methylene chloride extracts of ‘Snus’ are carcinogenic due to a genotoxic mechanism were both predicted to be low. Using carcinogenicity prediction by battery selection (CPBS), the probabilities of the methylene chloride and aqueous extracts being correctly identified as non-carcinogens are 71 and 77%, respectively. Up to date, the CPBS approach has been validated primarily for individual compounds, so some caution should at present be exercised in interpreting the results using this method. Based on these results, the carcinogenic potential of Swedish ‘Snus’ should be considered to be low, a conclusion in agreement with the low incidence of oral cancer in Sweden compared to other countries.

1,3-Butadiene and its epoxides induce sister-chromatid exchanges in human lymphocytes in vitro

Sister-chromatid exchanges (SCEs) were induced in human lymphocytes by 1,3-butadiene and its epoxides 3,4-epoxy-1-butene and 1,2:3,4-diepoxybutane. After a pulse treatment of 2 h, 1,3-butadiene produced a weak but reproducible increase in SCEs both with and without S9 mix. The response was similar in cultures of whole blood and of isolated lymphocytes. The 2 epoxide metabolites of butadiene, studied in whole-blood lymphocyte cultures without exogenous metabolic activation, were highly active SCE inducers. The lowest effective concentrations of butadiene, monoepoxybutene, and diepoxybutane were 2000 μM, 25 μM and 0.5 μM, respectively. A slight but dose-dependent increase in SCEs was also observed without an exogenous metabolic system after a 48-h treatment with 1,3-butadiene. Already the lowest concentration tested (500 μM) was effective. Again, the response was similar in cultures of whole blood and isolated lymphocytes, suggesting that the lymphocytes are capable of metabolically activating 1,3-butadiene.

Quantitative analyses of the induction of chromosome aberrations and sisterchromatid exchanges in human lymphocytes exposed to γ-rays and mitomycin-C in combination

Most chemicals are S-dependent and are potent inducers of SCE, but do not produce chromosome-type aberrations in the first metaphases after exposure. Ionizing radiation, which is an S-independent agent, produces chromosome-type aberrations, especially dicentrics and rings, but inefficiently produces chromatidtype aberrations. A series of experiments has been performed to investigate whether cytogenetic damage induced by ionizing radiation (γ-rays) might be assessed separately from that induced by the alkylating chemical, mitomycin C (MMC), when human lymphocytes were exposed to these 2 agents in combination. Whole-blood cultures of human lymphocytes in G o phase were exposed to γ,-rays and MMC in combination or separately. Cytogenetic analyses were done for both chromosome aberrations (CA), analyzed in cultures incubated for 56 h without BrdUrd, and sister-chromatid exchanges (SCEs) in cultures incubated for 72 h with BrdUrd. The frequency of chromosome-type aberrations (dicentrics and rings) increased with increasing doses of γ,-rays from 0.5 to 4.0 Gy. The dose-response relationships were the same with or without concomitant treatment with MMC (10 −6 M). Although the SCE frequency increased with increasing doses of MMC, the increase was nearly the same as when cells were treated with both MMC and γ-rays (2 Gy). There was no interaction between MMC and γ-rays concerning these 2 endpoints.

Genotoxic effects of thiram evaluated by sister-chromatid exchanges in human lymphocytes

The purpose of this investigation was to study the genotoxic potential of thiram (CAS No. 137-26-8) using an in vitro sister-chromatid exchange (SCE) assay with human lymphocytes. The results indicate that thiram and its metabolites increase the SCE frequencies 2-fold over those observed in the negative controls. The standard inducers cyclophosphamide and ethyl methanesulfonate increased SCE frequencies 10- and 4-fold, respectively, over untreated levels.

Hypotonic treatment leads to chromosomal aberrations but not to sister-chromatid exchanges in human lymphocytes

Human peripheral lymphocytes were isolated from whole blood and exposed to culture medium of reduced osmolality. This hypotonic treatment led to a significant increase in the frequencies of chromosomal aberrations when the osmolality was reduced to 60 mOsm/kg H 20 and below. Maximum damage occurred when the hypotonic treatment was done 27 or 30 h after starting the cultures. We also looked for the induction of sister-chromatid exchanges (SCE) by hypotonic culture conditions, but the SCE frequencies were not influenced.

Sister chromatid exchanges in human lymphocytes treated with silica.

The abilities of Min U Sil quartz or tridymite particles to induce sister chromatid exchanges (SCEs) in cultures of human lymphocytes plus monocytes or of human purified lymphocytes were investigated. With cultures of lymphocytes plus monocytes the level of SCEs was significantly enhanced after treatment with tridymite at the highest dose tested (50 micrograms/cm2). No effect was observed with purified lymphocytes. Quartz did not give clear cut results. Complementary experiments with tridymite filtrates suggested that phagocytosis of tridymite particles by monocytes was a necessary step for the induction of SCEs in human lymphocytes.

Sister chromatid exchange in lymphocytes of agricultural workers exposed to pesticides

Sister chromatid exchange (SCE) was studied in the lymphocytes of 27 agricultural workers occupationally exposed to several pesticides and 28 matched controls from el Maresme, an agricultural area near Barcelona. Comparison between both groups with the t-test did not reveal significant differences. These negative findings suggest that, possibly, the exposure level is too low to increase SCE in human lymphocytes in vivo. Our results indicate that smokers, both the workers and the controls, had a higher SCE frequency than non-smokers, in agreement with previous data reported by different authors.

Fecapentaene causes sister-chromatid exchanges in human lymphocytes

Fecapentaenes are potent mutagenic compounds found in human feces that are considered as potential colon carcinogens. It is demonstrated that a synthetic racemic all- trans fecapentaene-12 (fec-12) causes a strong dose-dependent increase in the frequency of sister-chromatid exchanges (SCE) in human lymphocytes exposed at different stages of the cell cycle. The SCE-inducing capacity is consistent with published results on the DNA-damaging activity of fec-12 such as formation of DNA single-strand breaks and interstrand cross-links.

Sister-chromatid exchanges in human lymphocytes exposed to 1- p-(3-methyltriazeno)benzoic acid potassium salt

The 1- p-(3-methyltriazeno) benzoic acid potassium salt (MTBA) is a triazeno analogue of dacarbazine, an antineoplastic agent capable of mediating the appearance of new antigenic specificities on cancer cells in mice, a phenomenon described as ‘chemical xenogenization’ (CX). Recently we reported the clastogenic potential of MTBA on human lymphocytes. Since sister-chromatid exchange (SCE) assay is more sensitive than clastogenic tests, at least at low drug concentrations, we assessed SCE frequencies induced by MTBA on human lymphocytes stimulated by PHA. Drug treatment at 2–500 μg/ml was performed in vitro prior to or after PHA addition. SCE values increased significantly in a dose-dependent manner up to 200 μg/ml. However, SCE frequencies, as well as chromosome breaks, did not increase dramatically. These data indicate that MTBA concentrations used for CX do not cause severe cytogenetic damage to immune cells at least in vitro.

Expression pattern of chromosomal aberrations and sister-chromatid exchanges in human lymphocytes

Expression pattern of chromosomal aberrations and sister-chromatid exchanges in human lymphocytes

Assessment of the mutagenic potential of a fungicide Bavistin using multiple assays

The fungicide Bavistin was assessed for mutagenic potential by various assays. Bavistin was found to be unable to induce gene mutation in Salmonella typhimurium, but it was able to induce transfection inhibition in Mycobacterium smegmatis. Bavistin was able to induce immediate genotoxic effects in plants but these were not carried through in development as in the long term no genotoxic effects were observed by the progeny test. Bavistin did induce micronuclei formation and did cause an increase in the ratio of normochromatic to polychromatic erythrocytes in mice. It was able to induce a very low frequency of sister-chromatid exchange in human lymphocytes and in addition, it was observed that the chemical affected the mitotic index but did not affect the cell cycle duration. Present studies indicate that the pesticide shows a positive response in 4 out of 5 different test systems (Table 8) and most of the observations support that Bavistin is genotoxic.

Doppler ultrasound pulses can induce SCE In human lymphocytes in vitro

Human lymphocytes were exposed to an ultrasound pulsing regimen similar to that typically employed in clinical Doppler measurements of fetal blood flow, to determine whether such pulses were able to alter the frequency of sister chromatid exchanges (SCE). A 3.1‐MHz, 1.5‐cm‐diam transducer emitted 5‐ to 50‐cycle pulses in a beam that was directed vertically along the axis of a stationary tube containing the blood suspension. Hydrophone measurements within the tube showed that the beam intensity profile was maintained beyond the free field focal region, thereby ensuring maximum exposure of the contents of the tube. A significant increase in SCE frequency was observed in blood from two random donors exposed to “diagnostic” levels of ultrasound, where the SPTA intensity ranged from 1 to 4 W/cm2. However, a number of further studies on blood from a single (different) donor have failed to show any effect from ultrasound even after an exposure dwell time of 24 h. This extraordinary finding may have some relevance towards explaining some of the controversial reports of SCE induction in human lymphocytes. Research continues to try to identify the mechanisms responsible for this effect.