Mutagenic activity of carcinogens and other chemical agents in Salmonella typhimurium tests

One-hundred and six chemical compounds were tested in Ames test with bacteria Salmonella typhimurium. Eight different strains (mainly, TA98 and TA100) were used. Liver S9 from Aroclor-treated rats was employed for metabolic activation. In group I comprising 51 compounds with sufficient evidence of carcinogenicity for animals, 45 were mutagenic while 6 (urethane, 1,2-dimethylhydrazine, DDT, chlorophorm, 1,4-dioxane and carbon tetrachloride) were not. In group 2 (27 noncarcinogenic compounds), 22 agents failed to exhibit mutagenicity whereas 5 (acrolein, styrene-oxide, acridine orange, I-naphthylamine and dichlormethane) revealed such activity. In groups 1 and 2, the sensitivity was 88.2, specificity--81.5 and predictive value--90%. In group 3 consisting of 28 agents used in chemical, pharmaceutical and food industries but not yet tested for mutagenicity and for the carcinogenicity of which no conclusive data are available, seven appeared to be mutagens (1-aminoanthraquinone, 2-aminoanthraquinone, 1-amino-4-chloranthraquinone, based blue, dinitrochlorbenzene, nitrosodiphenylamine and 2,3,5-trinitronaphthalene).     

Mutagenicity and carcinogenicity of fish sauce from a county with the high risk for gastric cancer in china

The mutagenicity and carcinogenicity of fish sauce (FS) sample from Changle County, a high gastric cancer mortality (113.20/10⁵) are, were investigated with the biologic short-term tests and laboratory animal experiment. The results showed that the extract of FS was markedly direct mutagenic towardS. typhimurium TA100, induced high sister chromatid exchanges (SCE) and micronucleus (MN) in V79 cells after nitrosation with sodium nitrite. But the non-nitrosated FS did not. The nitrosated fish sauce (NFS) also induced SOS inE. coli PQ37 and alkylation of calfthymus DNA. The potency of NFS to induce unscheduled DNA synthesis (UDS) in human normal gastric mucosal cells was increased about fivefold compared with FS. When the NFS extract was given to newborn rats by gavage, dysplasia and adenocarcinoma were induced in the glandular stomach in the 4th and 16th experimental week, respectively. N-nitrosamides were also found in NFS, which may account for the mutagenicity and carcinogenicity of NFS. It is indicated that FS, a traditional daily eaten seasoning, may contribute to the causes of the high gastric cancer mortality for the local residents.     

Metallothionein modulates the carcinogenicity of N-butyl-N-(4-hydroxybutyl)nitrosamine in mice.

We examined the carcinogenicity of N-butyl-N-(4-hydroxybutyl)nitrosamine (BBN) in transgenic mice deficient in the metallothionein (MT) I and II genes and in control (129/Sv) mice. Both strains of mice were given BBN for 8 weeks with or without Zn treatment. All mice were killed at 12 weeks after the cessation of BBN administration. BBN induced bladder tumors in 75% of MT null mice and in 43% of 129/Sv mice. The average number of bladder tumors per mouse was significantly higher in MT null mice (1. 18 +/- 0.27) than in 129/Sv mice (0.43 +/- 0.20). Zn treatment suppressed the carcinogenicity of BBN in 129/Sv mice but not in MT null mice. Histopathological examination of the tumors revealed that the malignant potential of bladder tumors in 129/Sv mice was greater than that in MT null mice. These results indicate that MT is an important modulator of carcinogenicity of BBN in the bladder of mice.     

Mutagenicity of pyrolysates of salt-tolerant bacteria from food and cigarettes

Smoke condensates obtained by pyrolysis of the cells of microorganisms isolated from food and cigarettes and of laboratory cultures were shown to be mutagenic to Salmonella typhimurium TA98 and TA100 in incorporation with liver microsomal fraction S-9 mix. The smoke condensates from salt-tolerant bacteria, which belonged to Micrococcus, Bacillus and Corynebacterium spp., showed higher mutagenic activity than those from other less salt-tolerant organisms, fish flesh or pork. Smoke deriving from microorganisms, especially salt-tolerant ones, in food or cigarettes as well as from food components might take part in human carcinogenesis because of the correlation between mutagenicity and carcinogenicity.     

Quantitative comparisons between carcinogenicity, mutagenicity and electrophilicity of direct-acting N-nitroso compounds and other alkylating agents

The quantitative relationship between carcinogenicity in rodents and mutagenicity in S. typhimurium was examined using 10 monofunctional alkylating agents, including N-nitrosamides, alkylmethane sulfonates and epoxides. The compounds were assayed for mutagenicity in two S. typhimurium strains (TA1535, TA100) and in plate and liquid assays. Mutagenic activity was compared with alkylating activity, half-life (solvolysis constant) and carcinogenic activity in rodents (TD50). No correlations between these variables were found. However, there was a good positive relationship between the TD50 values of the carcinogens and the initial ratios of N-7-alkylguanine to 0(6)-alkylguanine formed after reaction with double-stranded DNA in vitro (r = 0.88, p less than 0.01; n = 9). From these results, it is concluded that the N-7/0(6)-alkylguanine ratio is quantitatively related to the carcinogenic activity for this class of compounds and it may therefore be used to predict the carcinogenic potency of new compounds within this class.     

Mammary carcinogenesis and molecular analysis of in vivo cII gene mutations in the mammary tissue of female transgenic rats treated with the environmental pollutant 6-nitrochrysene

We determined the mutant fractions (MF) and mutational specificities in the cII gene in histologically confirmed normal, non-involved and tumor mammary tissues of female transgenic (Big Blue F344 x Sprague-Dawley)F1 rats treated with the environmental pollutant 6-nitrochrysene (6-NC). At 30 days of age, three groups were set up for oral treatment with 6-NC dissolved in trioctanoin, or trioctanoin alone once a week for 8 weeks. Two dose levels of 6-NC (100 and 200 micromol/rat) were selected on the basis of our previous carcinogenicity bioassays with CD rats. The rats were decapitated 32 weeks after the last carcinogen dose. Both incidence and multiplicity of mammary adenocarcinomas were significantly elevated in the high dose (36%, 0.57, P < 0.01) group but at the low dose these outcomes (16%, 0.23, P < 0.1) were not significantly different from those of control rats (3%, 0.03). The MF in normal, non-involved and tumor tissues from the mammary glands of 6-NC-treated rats were comparable. At the high and low doses, respectively (4.8 +/- 2.0, 3.2 +/- 2.1) the MF of 6-NC-treated rats, were significantly higher (P < 0.05) than that observed in control rats (1.2 +/- 0.6). Control mutants consisted primarily of GC --> AT transitions, whereas 6-NC-induced mutants were comprised of several major classes of mutations with GC --> TA, GC --> CG, AT --> GC and AT --> TA as the most prevalent. Further studies indicated that the structures of 6-NC-DNA adducts in the mammary tissue are consistent with the mutational specificities. This is the first report that defines the relationship between carcinogenesis and mutagenesis, as well as between structures of 6-NC-DNA adducts and mutation characteristics in the target organ in vivo.     

Effect of gefitinib on N-nitrosamine-4-(methylnitrosamino)-1-(3-pyridyl)-1-butanone induced lung tumorigenesis in A/J mice

Gefitinib is an epidermal growth factor receptor tyrosine kinase inhibitor (EGFR-TKI). N-Nitrosamine-4-(methylnitrosamino)-1-(3-pyridyl)-1-butanone (NNK), a potent carcinogen found in tobacco smoke, induces lung tumors in A/J mice. NNK induces cellular transformation resulting in the over-expression of EGFR. Accordingly, EGFR may be a target for cancer prevention. In this study, we investigated the effect of gefitinib on NNK-induced tumorigenesis and the carcinogenicity of gefitinib in A/J mice.A total of 180 four-week-old female A/J mice were randomly divided into six groups: group 1 (controls), treated with deionized water; group 2, treated with 5 mg/kg p.o. gefitinib; group 3, treated with 50 mg/kg p.o. gefitinib (to test the carcinogenicity of gefitinib); group 4 (controls for NNK treatment), treated with deionized water; group 5, treated with 5 mg/kg p.o. gefitinib; and group 6, treated with 50 mg/kg p.o. gefitinib and injected with NNK once at 8 weeks of age to test the chemopreventive activity of gefitinib. Gefitinib was given once a day, 5 days a week by gavage, beginning at 4 weeks of age and continuing for 26 weeks. All mice were sacrificed at 30 weeks of age. The multiplicities of the NNK-induced lung tumors were significantly suppressed in a dose-dependent manner. Gefitinib had no effect on body weight at a low dose. The administration of gefitinib alone for 26 weeks did not induce tumorigenesis; instead, it significantly suppressed the incidence of spontaneous tumors in the mice, in contrast with other anti-cancer agents. Gefitinib did not induce lung fibrosis when compared with control mice by Azan–Mallory staining. Our results suggest that gefitinib has a weak but significant chemopreventive effect with no carcinogenicity or pulmonary toxicity in A/J mice.     

Incidence of nonmelanoma skin cancer in the United States: J. Scotto,T.R. Fears and J.F. Fraumeni,U.S. Dept. Health and Human Services,Public Health Service,National Institute of Health,National Cancer Institute. NIH Publication No. 83–2433 (1983)

Two hydrazine derivatives, monoacetyl hydrazine (MAH) and diacetyl hydrazine (DAH), have been tested for mutagenic response in the Salmonella/mammalian microsome assay and micronucleus test. MAH but not DAH, increased the revertant mutants in TA100 and TA1535 and also increased the frequency of micronuclei in polychromatic erythrocytes. Gavage administration of MAH but not of DAH, resulted in increased incidence of lung tumors. These observations record for the first time the mutagenicity/carcinogenicity of MAH which is one of the metabolites of isoniazid in animals and humans.     

A quantitative structure-activity analysis of the mutagenic and carcinogenic action of 43 structurally related heterocyclic compounds

Quantitative structure-activity relationships for the carcinogenicityand mutagenicity (against Salmonella typhimurium his- TA98,TA100 and TA1537 strains) of 43 structurally related heterocycliccompounds were formulated. The compounds investigated belongto the following two series of congeners: (a) benzo(thio)-pyranoquinolinesand (b) benzo(thio)pyranoindoles. Their biologic activitieswere correlated with the following parameters: (a) the minimaltopological difference (describing the fit of the consideredmolecules with a possible receptor, enzyme or DNA) and (b) thelipophilicity constants. The computed regression equations suggestthat the structural requirements for carcinogenicity are differentfrom those responsible for mutagenicity in the Ames test.     

Effect of dietary protein on the response of rainbow trout (Salmo gairdneri) to aflatoxin B1.

Diets containing either 49.5% or 32% casein or fish protein concentrate (FPC) were fed to young rainbow trout (Salmo gairdneri) for 12 months. Five levels [0, 2, 6, 18, and 54 parts per billion (ppb)] of aflatoxin B1 (AFB1) were given in each of four different diets. A 30-fish sample was taken at 6, 9, and 12 months to determine the influence of diet on the carcinogenicity of AFB1. Both levels of casein produced similar hepatoma incidences at each level of AFB1. The diet high in FPC produced more tumours than did the casein diets at 2, 6, and 18 ppb AFB1, whereas fish fed the diet low in FPC had a significantly (P less than 0.05) lower hepatoma incidence than did the other three groups. The liver size (percent body wt) was smaller at higher toxin levels in all instances. The growth of fish given 32% casein was less than that of the other groups.     

Mutagenicity and carcinogenicity of smoked meat from Nagaland, a region of India prone to a high incidence of nasopharyngeal cancer

The incidence of nasopharyngeal cancer (NPC) in the north-easternpart of India is reported to be high. A possible correlationbetween consumption of smoked meat by the tribal people andhigh suceptibility to NPC has been postulated. The charred portionof smoked beef and meat of other animals was collected fromthis area, extracted with acetone and the extract (SME) wastested using the Ames test as well as for chromosomal aberrationin mouse bone marrow cells and carcinogenicity using Swiss baremice. It was oberved that SME was mutagenic in all five strainsof Salmonella typhimurium (TA98, TA1538, TA100, TA1535 and TA1537),with or without S9 mix, and was clastogenic in a mammalian testsystem. SME also has the potential to induce skin papillomaas well as systemic tumours in Swiss bare mice. Chemical analysisof SME revealed the presence of low concentrations of volatilenitrosamines.     

The reduced mutagenicity of aflatoxin B1 due to hydroxylation: observations on five Salmonella typhimurium tester strains.

The mutagenicity of aflatoxin M1 relative to that of aflatoxin B1, the parent compound, was studied in 5 Ames' tester strains of Salmonella typhimurium (TA 98, TA 100, TA 1535, TA 1537, TA 1538). Aflatoxins B1 and M1 are both highly mutagenic in microsome-mediated system in TA 100. The prediction of the relative carcinogenicity of aflatoxin M1 to aflatoxin B1 posed by the mutation of TA 100 is probably more authentic than by TA 98.     

Effects of the carcinogen, acrylonitrile, on forestomach cell proliferation and apoptosis in the rat: comparison with methacrylonitrile

Acrylonitrile (AN) and methacrylonitrile (MAN) are two major industrial nitriles used in the production of plastics and acrylic fibers. Whereas AN is a potent acute toxin and carcinogenic in rats, little is known regarding MAN. Current work is part of an overall effort designed to assess the potential toxicity/carcinogenicity of MAN. The present study compares the ability of the two chemicals to induce epithelial proliferation and apoptosis in the forestomach (FS; a target of AN carcinogenicity), liver and glandular stomach (non-targets of AN carcinogenicity) of male F344 rats. AN was administered to rats daily, by gavage, for 6 weeks, at 0.43 and 0.22 mmol/kg. MAN was administered at 0.87 and 0.43 mmol/kg. Both AN and MAN induced a dose-dependent increase in epithelial cell proliferation in the FS of male F344 rats as determined by bromodeoxyuridine (BrdU) incorporation into DNA. In contrast, AN, but not MAN caused a dose-dependent increase in the thickness of the forestomach squamous mucosa. This increased thickness (hyperplasia) was reflected by an increase in the number of total epithelial cells per unit length of mucosa. At doses of AN and MAN which induced a 2.3-fold increase in BrdU incorporation, apoptosis was 5- and 18-fold greater than controls, respectively. Although both MAN and AN caused a similar increase in cell proliferation, the relatively more prominent increase in the apoptotic index of the squamous epithelium of rats exposed to MAN may explain the lack of a detectable increase in the thickness of the mucosa compared to that seen with AN. The disruption of the balance between FS mucosal cell proliferation and apoptosis in favor of a net increase in the number of FS epithelial cells per unit length may contribute to the carcinogenicity of AN. In conclusion, present work demonstrated that AN selectively induced a net enhancement in FS cell proliferation, a site of its carcinogenicity. On the other hand, MAN-induced FS cell proliferation was associated with a parallel increase in apoptosis. The relatively greater increase in apoptosis by MAN may have compensated for the increase in FS mucosal cell proliferation and the lack of observable change in the FS thickness.      

Plant tannins as inhibitors of hydroperoxide production and tumor promotion induced by ultraviolet B radiation in mouse skin in vivo.

The anti-oxidant and anti-tumor promotion activities of several tannins extracted from plants were examined in mouse skin treated with ultraviolet B (UVB) radiation in vivo. Hydroperoxide production was found to be maximally stimulated at a UVB dose of 200 mj/cm2, beyond which no further stimulation occurred. Treatment of mouse skin with two UVB doses of 225 mj/cm2 each, applied at a 48 h interval gradually increases the hydroperoxide (HPx)-producing activity of the epidermis, which is maximally stimulated at 4 days and returns to control levels at 15 days. The magnitude of the HPx response is found to increase with repeated UVB treatments applied at a 48 h interval and reaches a maximum level following four treatments. Of the three tannins tested (Commercial TA, Tarapod TA, and Oak TA), Tarapod TA is found to be the most effective inhibitor of UVB-stimulated HPx activity. Pretreatment with Tarapod TA inhibits, in a dose-dependent manner, this HPx response to UVB radiation. Inhibition by Tarapod TA occurs when it is applied at distant times before (-12 h) or after (+24 h) UVB radiation. When applied 20 min before UVB radiation, twice a week for 25 weeks, 8 mg of Tarapod TA inhibits the incidence and yield of papillomas promoted by UVB light in initiated skin by 34 and 70% respectively. Furthermore, when 10 mg/kg of mouse body weight of Tarapod TA was injected intraperitoneally, for a period of 25 weeks, 20 min prior to UVB treatment, it inhibited the yield of papillomas by 44%, suggesting that plant tannins when administered by various means are useful photoprotectants.     

Neoplasia and chronic disease associated with the prolonged administration of dehydroheliotridine to rats

When dehydroheliotridine (DHH), a pyrrolizidine alkaloid metabolite with bifunctional alkylating and antimitotic activities, was administered to a hooded strain of rats by ip injection, the incidence of tumors, excluding interstitial cell tumors, was significantly greater than that in saline-injected controls. The number of tumors was not further increased when thioacetamide (TA) was co-administered for its mitosis-stimulating effect. The life-span of the rats was significantly shortened by DHH and more so by combined DHH and TA treatment, but not by TA alone. The results indicate that DHH is responsible for some, possibly most, of the carcinogenicity of the parent pyrrolizidine alkaloids and also stimulates the earlier and more rapid development of renal and vascular diseases normally associated with aging in rats.     

Induction of lung tumors and lymphomas in BALB/c mice by metronidazole

Metronidazole, which is widely used in the treatment of "Trichomonas vaginalis", "Entamoeba histolytica" and "Giardia lamblia" infections, was administered to BALB/c mice by stomach tube in an aqueous solution at a dose rate of 2 mg/day for 100 days (total 200 mg) to test its carcinogenicity. The treatment induced a significant increase in lung tumors in male mice (p less than 0.001) and provoked the appearance of lymphomas in females (p less than 0.001). Although there is insufficient evidence to pass judgement on the potential carcinogenicity of metronidazole in man, the results of this and other investigations reported in the literature have demonstrated that metronidazole develops carcinogenic activity in rats and mice.     

Carcinogenesis in rat esophagus by intraperitoneal injection of different doses of methyl-n-amylnitrosamine.

The carcinogenicity of methyl-n-amylnitrosamine in MRC-Wistar rats was determined after i.p. injection at a variety of dose schedules. After 6 weekly methyl-n-amylnitrosamine injections of 25 mg/kg or 12 weekly injections of either 12.5 or 25 mg/kg, the incidence of esophageal squamous cell papillomas was 85 to 100% and that of esophageal squamous cell carcinomas was 40 to 65%. With 12 injections, the mean survival time was 25 to 31 weeks. Treatment with 1 or 2 doses of 50 mg/kg produced a lesser incidence (less than 20%) of esophageal tumors, with a longer survival time of 67 to 77 weeks. One 85-mg/kg injection caused esophageal carcinomas in 5 of 7 rats. The treated groups also had squamous cell papillomas and carcinomas in the nasal cavity (up to 50% incidence) and trachea (up to 30% incidence). Hence, a 6- or 12-week treatment schedule was adequate for inducing esophageal tumors and could be used for studies on agents modifying esophageal tumor induction by methyl-n-amylnitrosamine.     

Experimental study on the carcinogenicity of the Cytostatic drug ftorafur (tegafur)

Long-term carcinogenicity of Ftorafur (Tegafur) was studied in rodents. Rats and mice were treated for one year per os with 40 (mice) and 60 (rat) mg/kg Ftorafur twice a week and were followed for their entire life. Analysis of the data provide no evidence for the carcinogenicity of Ftorafur in rodents. These findings are similar to other antimetabolite studies and contrasts with the carcinogenic alkylating agents.     

Mutagenicities of smoke condensates and the charred surface of fish and meat.

Smoke condensates obtained from broiling fish showed mutagenic activity for Salmonella typhimurium TA100 and TA98. Metabolic activation was required to induce mutagenic activity of smoke condensates of some species of fish. The smoke condensate obtained during charcoal broiling of beefsteak was far less mutagenic than that of fish, with or without metabolic activation. Extracts of the charred surface of broiled fish and meat also contained mutagenic substances. These extracts needed metabolic activation to exhibit mutagenicities on TA98. The mutagenic activity of the smoke condensate obtained from one sardine weighing 100 g was equivalent to that of 132 micrograms benzo(a)pyrene and that of the charred surface of the sardine was equivalent to 358 micrograms benzo(a)pyrene. One piece of beefsteak weighing 190 g, contained mutagenic activity equivalent to that of 855 micrograns benzo(a)pyrene.