Mutation Research/Genetic Toxicology最新文献

We examined the binding of various steroid hormones to DNA in vitro by means of ³²P-postlabeling. Seventeen steroid hormones and cholesterol (CS) were incubated with human liver DNA at 37°C for 1 h under aerobic conditions in the absence of catalysis. The reaction mixtures were analyzed by the nuclease P-1 version of ³²P-postlabeling. The results showed that cortexolone (CX), prednisolone (PS), cortisone (CN), cortisol (CL), tetrahydrocortisol (TC), corticosterone (CC), 11-deoxycorticosterone (DC), dexamethasone (DX), dihydrocortisol (DL), and aldosterone (AL) covalently bound with DNA. However, progesterone (PG), 17α-hydroxyprogesterone (HG), estrone (E1), estradiol (E2), estriol (E3), testosterone (TS), cortol (CR) and the original compound for biosynthesis, CS, did not form adducts. In absence of DNA, the steroids themselves did not give rise to any spot on TLC under the same conditions. The dose-responses of DNA binding by DC, DL, CC, CL and CN were linear. The relative adduct labeling of reactive steroids at a concentration of 2 mM were as follows: 68.8 (CX), 53.2 (PS), 39.6 (CN), 29.9 (CL), 20.9 (TC), 12.9 (CC), 12.3 (DC), 7.5 (DX), 4.7 (DL), 1.2 (AL) adducts per 10⁸ nucleotides. Reactive and nonreactive steroids were distinguishable by the presence or absence of the carbonyl group (-CO-CH₂OH) at carbon seventeen (C₁₇) of the cholesterol skeleton. This implies that the electrophilic carbonyl or a neighboring group perhaps involved in the formation of covalent bond with DNA. To investigate the nature of target base(s) of these DNA reactive steroids, mononucleotides of all four bases of DNA were reacted with CN, CL, CC and cochromatographed with the obtained spots of DNA reactions. The results of which stated that these steroids and guanine reaction gave the same spots as observed in DNA reaction, indicating guanine is the main target of these DNA reactive steroids. Hep G2 human hepatocellular carcinoma cells were used as an alternative model. Although nine steroids (CL, DL, TC, PS, DX, PG, E2, TS, CR) did not react with intracellular DNA under our experimental conditions, our findings suggested that some hormonal steroids can form covalent DNA adducts in vitro.

Chromosomal aberrations (CA) were studied in the peripheral blood lymphocytes of 85 healthy male volunteers from Barcelona (Catalonia, Spain). The effect that factors such as age, life style, work exposure and medical treatment had on the cytological endpoints was studied by means of a Poisson regression model. The results obtained indicate a significant positive relationship between the age of the subjects analyzed and the total of chromosome-type aberrations. With respect to the other variables analyzed, a positive association was found between the frequent consumption of analgesics and the incidence of chromatid-type aberrations.

In the absence of a metabolizing system (S9 mix) 4-chloro-o-toluidine (4-COT) was found to be ineffective in a combination of assays for gene mutations in Salmonella typhimurium, for chromosome aberrations and sister chromatide exchanges in human lymphocytes, and for the induction of spindle disturbances in V79 Chinese hamster cells. In the presence of S9, 4-COT was also ineffective in producing structural or numerical changes in mammalian cells, but the yields of 4-COT induced revertants in S. typhimurium strains TA 100 and TA 98 were about 2-fold higher than those in controls.

Electroplating effluents were tested for their genotoxicity with the micronucleus test on newt larvae. The metallic content of the tested samples was responsible for the induction of micronuclei in red blood cells (RBC). Then, iron (Fe³⁺), chromium (Cr³⁺, Cr⁶⁺) and zinc (Zn²⁺) which were identified in these samples, were tested either separately or combined, at their concentrations in the electroplating effluents. Fe³⁺ induced a high level of micronuclei at 12.5 and 25 mg/l (nominal concentrations). Both soluble and non-soluble forms of iron were responsible for these genotoxic effects. At lower concentrations (0.6 and 4.5 mg/l) Fe³⁺ was not systematically genotoxic. Zinc could not be considered genotoxic on newt. Cr³⁺ gave negative responses, but exposure to Cr⁶⁺ (1 mg/l) could result in a significant number of micronucleated RBC in some cases. The most dramatic genotoxic effects were registered when Fe³⁺ and Cr⁶⁺ were combined. This study demonstrates that interactions between pollutants and the effects of non-soluble chemicals on aquatic vertebrates and invertebrates can no longer be neglected.

The potentially protective role of chlorophyllin, the sodium and copper salt of chlorophyll a against the initiation and promotion stages in carcinogenesis was studied by in vitro short-term assays. Chlorophyllin showed a dose-dependent suppressive effect on 3-amino-1,4-dimethyl-5H-pyrido[4,3-b]indol (Trp-P-1)-induced umu C gene expression of Salmonella typhimurium (TA 1535/pSK 1002) in the presence of metabolizing enzyme mixture. The similar inhibitory effect of chlorophyllin was detected in mitomycin C (MMC)-dependent umu C gene expression in the absence of metabolizing enzyme mixture. Furthermore chlorophyllin also exhibited a dose-dependent inhibition on 12-O-tetradecanoyl-phorbol-13-acetate (TPA)-induced ornithine decarboxylase (ODC) activity of 3T3 fibroblast cells at the same concentrations. However, when chlorophyll a isolated from Japanese tea leaves was applied on the same assay systems as a comparative experiment, chlorophyll a showed much weaker activity compared with that of chlorophyllin. The significance of this finding is discussed from the viewpoint of the protective role of chlorophyllin against carcinogenesis.

The Loats Automated Micronucleus Scoring System was developed to assist with the evaluation of compounds for the ability to induce micronuclei in mouse bone marrow polychromatic erythrocytes (PCE). This image analysis system can identify PCE as well as normochromatic erythocytes (NCE) and calculate the PCE/NCE ratio as an index for bone marrow toxicity. Two studies were conducted to provide slides for a comparison of micronucleated PCE values collected manually to those collected by the automated system. Mitomycin C was used as a micronucleus-inducing agent to elicit a positive response and Lilly compound 303497 was used as an example of a compound negative for the induction of micronuclei. No statistically significant differences were observed between micronucleus counts obtained manually and those obtained by the automated system. The PCE/NCE ratios calculated by the automated system were also similar to those determined from the manually collected PCE and NCE counts for the vehicle and positive controls, however, differences in the ratios were observed in compound treatment groups. These differences were attributed to a larger population of transitional cells in the treated groups. These results confirm that the Loats Automated Micronucleus Scoring System is an acceptable alternative to manual evaluation of mouse bone marrow slides for the incidence of micronucleated PCE.

In vivo sister chromatid exchange (SCE) and chromosome aberrations (CA) were carried out for six salicylic acid derivatives in bone marrow cells of mice. Six salicylic acid derivatives, namely acetyl salicylic acid (aspirin), salicylic acid, salicylamide, sodium salicylate, diflunisal and niclosamide, were used for these experiments. Drugs were administered both intraperitoneally (i.p.) and orally by gavage. Out of these six salicylic acid derivatives tested, only diflunisal and niclosamide showed genotoxicity as measured by both SCE and CA assays. Acetyl salicylic acid and sodium salicylate showed weak genotoxicity as measured by SCE and CA, respectively, only at the highest dose tested.

The protective effect of β-carotene (β-C) and α-tocopherol (α-T), singularly and in equimolar mixtures, toward the photomutagenicity induced by 8-methoxypsoralen (8-MOP), at different oxygen partial pressure (pO₂), was evaluated in two different experimental models: Salmonella typhimurium TA102 and Saccharomyces cerevisiae D7. After phototreatment with 8-MOP, the results show a lethal effect under hypoxic conditions in both experimental model systems, an increase in revertants associated to the pO₂ increase in S. typhimurium TA102, and a decrease in revertants and convertants associated to the pO₂ increase in S. cerevisiae D7. In S. typhimurium TA102, in atmospheric condition, β-C and α-T (1.86 or 18.6 μM) show a protective effect only at the higher dosage. α-T was more protective than β-C. The equimolar mixtures show an antimutagenic effect at both dosage used with a synergistic effect at lower dosage and an additive antimutagenic activity at higher dosage. An inhibition of the spontaneous mutagenicity by mixtures at higher dosage was also observed. The results obtained in S. typhimurium TA102 show an antimutagenic effects of β-C, α-T and their mixture at 190 mmHg pO₂, confirming the data obtained in air condition. At 380 mmHg pO₂, α-T and the mixture show a significant antimutagenic activity; at 570 mmHg pO₂, only α-T is protective. At 760 mmHg pO₂, no protective effect was observed by the two antioxidants, and β-C increases the photomutagenicity induced by 8-MOP. In S. cerevisiae D7 a protective effect was only observed at 380 mmHg pO₂ with the mixture. No antigenotoxic effect was found in the other experimental conditions, even if the uptake of the two antioxidants was confirmed by HPLC. Our results underline the role of oxygen in the photomutagenicity induced by 8-MOP and in the antimutagenic activity of β-C and α-T. This is the first report confirming in a cellular experimental model the data obtained in some chemical systems: the protective effect of β-C only at low pO₂ and the synergistic effect of mixture of β-C and α-T.

The metabolites of 2-nitrofluorene (NF), 5-, 7- and 9-OH-2-nitrofluorene (OH-NF) were compared for their genotoxicity. Seventy-two hours after intraperitoneal administration of these substances individually to rats (100 mg/kg body wt.), DNA adducts in liver tissue were analyzed with ³²P-TLC and ³²P-HPLC. An in vivo liver model was used to test the initiating capacity of the said substances for the formation of preneoplastic lesions. 5-OH-NF showed low capacity to induced DNA adduct formation and low potential as initiator to induce preneoplastic lesions-foci/nodules in the liver of rats. Both 7- and 9-OH-NF induced DNA adducts and preneoplastic liver lesions but with smaller quantities compared to NF. It seems that 7- and 9-OH-NF can not be considered as detoxification products of NF. In general, the initiating capacity of these substances for the formation of preneoplastic lesions has a good correlation with their potency to form DNA adducts.