Tumors In B6C3F1 Mice Research Articles

Induced Cytotoxicity and Cell Proliferation in the Hepatocarcinogenicity of Chloroform in Female B6C3F1 Mice: Comparison of Administration by Gavage in Corn Oil vs ad Libitum in Drinking Water

Induced Cytotoxicity and Cell Proliferation in the Hepatocarcinogenicity of Chloroform in Female B6C3F1 Mice: Comparison of Administration by Gavage in Corn Oil vs ad Libitum in Drinking Water. Larson, J. L., Wolf, D. C., and Butterworth, B. E. (1994). Fundam. Appl. Toxicol. 22, 90-102.Chloroform increases the incidence of liver tumors in B6C3F1 mice when administered in by gavage in corn oil, but not when given in the drinking water at similar daily doses. Since cytotoxicity and regenerative cell proliferation have been implicated in the tumorigenic process for this nongenotoxic agent, these effects of chloroform in corn oil and drinking water were evaluated under conditions similar to the two bioassays. Female B6C3F1 mice were administered oral doses of 0, 3, 10, 34, 90, 238, or 477 mg/kg chloroform dissolved in corn oil 5 days/week for periods of 4 days or 3 weeks, or were continually exposed to chloroform in the drinking water at concentrations of 0, 60, 200, 400, 900, or 1800 ppm for 4 days or 3 weeks, at which time they were necropsied. 5-Bromo-2′-deoxyuridine (BrdU) was delivered via osmotic pumps implanted 3.5 days prior to necropsy. Cell proliferation was evaluated as the percentage of hepatocytes that entered S-phase over 3.5 days (labeling index, LI), measured by immunohistochemical detection of BrdU incorporated into the DNA. Dose-dependent changes included centrilobular necrosis and markedly elevated LI in mice given 238 or 477 mg/kg chloroform in corn oil (the average daily doses that produced tumors in the cancer bioassay). The no-observed-effect level for histopathological changes was 10 mg/kg/day and for induced cell proliferation was 34 mg/kg/day for chloroform given in corn oil. Chloroform given in the drinking water did not increase the hepatic LI after either 4 days or 3 weeks in any of the dose groups, nor were any microscopic alterations observed in the livers, even though the cumulative daily amount of chloroform ingested in the 1800-ppm exposure group was 329 mg/kg/day. The sustained increase in LI in the livers of mice administered hepatocarcinogenic doses of chloroform in corn oil, but not for chloroform in drinking water, is evidence that chloroform-induced mouse liver cancer is secondary to events associated with induced cytolethality and cell proliferation. The triggering of these effects appears to be dependent on both the rate and duration of chloroform delivery to the target tissues. Thus, the most straightforward risk assessment for chloroform for this tissue would assign no increased cancer risk for dosing regimens that do not induce cytolethality and cell proliferation.

In Vitro Cytotoxicity of Mono-, Di-, and Trichloroacetate and Its Modulation by Hepatic Peroxisome Proliferation

In Vitro Cytotoxicity of Mono-, Di-, and Trichloroacetate and Modulation by Hepatic Peroxisome Proliferation. Bruschi, S. A., and Bull, R. J. (1993). Fundam. Appl. Toxicol. 21, 366-375.Dichloroacetate (DCA) and trichloroacetate (TCA) are major by-products of drinking water chlorination. Recent experiments have shown that both of these compounds produce hepatic tumors in B6C3F1 mice. There was evidence that these effects may be associated with cytotoxic effects and/or peroxisomal proliferation. Therefore, in the present study the in vitro cytotoxicity of monochloroacetate (MCA), DCA, TCA and a metabolite, glycolate (GLY), was determined in hepatocyte suspensions prepared from naive and clofibric acid-pretreated male Sprague-Dawley rats and B6C3F1 mice. Cytotoxic responses, measured by release of lactic dehydrogenase and/or trypan blue exclusion, were only observed with high concentrations (5.0 mM) of MCA and GLY in hepatocytes from naive animals (p = 0.025 and 0.008, respectively, Sprague-Dawley rat; p = 0.033 and 0.001, respectively, B6C3F1 mouse). The cytotoxic responses to both compounds were observed much earlier and at much lower concentrations in hepatocytes taken from mice and rats that had been pretreated with clofibric acid (p ⩽ 0.001, Sprague-Dawley rat and B6C3F1 mouse). DCA and TCA produced no evidence of cytotoxicity in hepatocytes from naive or clofibric acid-pretreated animals of either species at concentrations up to 5.0 mM. Increasing concentrations of MCA and GLY resulted in dose-related depletion of intracellular reduced glutathione (GSH) that closely paralleled the cytotoxic responses. Only GLY (0.25-5.0 mM) produced increased intracellular oxidized glutathione. Neither DCA nor TCA was found to alter cellular GSH status in hepatocytes isolated from either Sprague-Dawley rats or B6C3F1 mice. It was concluded from these in vitro observations that DCA and TCA are not highly cytotoxic to hepatocytes. Moreover, the rates of their conversion to MCA or GLY may be insufficient to induce cytotoxic effects in hepatocytes in vivo.

The role of dichloroacetate in the hepatocarcinogenicity of trichloroethylene

The induction of hepatic tumors in B6C3F1 mice treated with trichloroethylene (TRI) has been attributed to its metabolism to trichloroacetate (TCA). Trichloroacetate is an effective peroxisome proliferator in mice at blood concentrations that are readily achieved with carcinogenic doses of TRI. Recent data has demonstrated that both TCA and diehloroacetate (DCA) are capable of inducing liver tumors in B6C3F1 mice. Although long recognized as a metabolite of TRI, little attention has focussed on the role DCA might play in the hepatocarcinogenic effects of TRI. There are significant differences in the effects of DCA and TCA on the liver of B6C3F1 mice. Trichloroacetate treatment induces peroxisome proliferation, increases lipid deposition, and results in a marked accumulation of lipofuscin in the liver with long-term exposures. Dichloroacetate induces a markedly enlarged liver associated with a cytomegaly and large accumulations of glycogen. The cytomegaly is associated with the development of focal areas of recurrent liver necrosis which in turn lead to high levels of cell proliferation in the area surrounding these lesions. Induction of peroxisomes with DCA is transitory and the accumulation of lipofuscin is much less evident than with TCA treatment. Studies of TRI metabolism demonstrate that blood levels of DCA produced are sufficient to account for the hepatocarcinogenic effects of TRI. The rather low concentrations of DCA found in the urine of mice treated with TRI relative to TCA concentrations are due to the much more rapid and complete metabolism of DCA. These data do not support the conclusion that the hepatocareinogenic effects of TRI are simply related to peroxisome proliferation.

Spontaneous neoplasms in B6C3F1 mice

Spontaneous neoplasms in untreated B6C3F1 mice (200 males and 200 females) used as controls in 4 carcinogenicity studies were evaluated and tabulated. The most common neoplasms in male mice were hepatocellular adenomas/carcinomas (24.5%) followed by alveolar-bronchiolar adenomas/carcinomas (10.0%), lymphoreticular neoplasms (7.0%) [malignant lymphomas mixed (4.5%), histiocytic sarcomas (3.5%)], harderian gland adenoma (6.5%), and hemangiomas/hemangiosarcomas (5.5%). In the females, the most frequently occurring neoplasms were lymphoreticular neoplasms (22.0%) [malignant lymphoma mixed (10.0%), malignant lymphoma lymphocytic (6.5%), histiocytic sarcomas (5.5%)] followed by pituitary adenomas (15.5%), alveolar-bronchial adenomas/carcinomas (11.5%), hepatocellular adenomas/ carcinomas (7.0%), harderian gland adenomas, uterine stromal polyps (2.5%), and hemangiomas/hemangiosarcoms (2.0%). The incidence of tumors in various other organs was found to be low.

Histomorphologic Features of Spontaneous and Chemically-Induced Pulmonary Neoplasms in B6C3F1 Mice and Fischer 344 Rats

The histomorphologic features of spontaneous and chemically-induced lung neoplasms in male and female B6C3F1 mice and Fischer 344 rats are described. Primary pulmonary neoplasms in mice and rats were classified as alveolar/bronchiolar (A/B) adenoma or carcinoma (including variants with squamous and mucinous cell differentiation), bronchial adenoma or carcinoma, squamous cell carcinoma or mesenchymal tumors. A/B adenomas and carcinomas were the most common spontaneous pulmonary neoplasms observed in both mice and rats, but were observed less frequently in rats. In the National Toxicology Program (NTP) historical control database the incidence of spontaneous A/B adenomas in male (n = 2,084) and female (n = 2,079) mice is 13.8% and 4.9%, respectively; for A/B carcinomas, it is 5.3% and 2.4%, respectively. In male (n = 3,877) and female (n = 3,919) rats, spontaneous pulmonary neoplasms are rare with historical control rates less than 3% for A/B adenomas or carcinomas in either sex. The spontaneous A/B adenomas and carcinomas observed in mice and rats typically had papillary, solid or mixed (papillary and solid) histologic growth patterns. Pulmonary neoplasms from mice and rats treated with chemical carcinogens reviewed from 2-year studies consisted primarily of A/B adenomas and carcinomas. These tumors had papillary, glandular/tubular, solid or mixed (combination of 2 or more) histologic growth patterns. A few of the A/B neoplasms had areas of squamous or mucinous cell differentiation. Other less frequently occurring spontaneous and chemically-induced neoplasms included squamous cell carcinomas, bronchial adenomas and carcinomas, and sarcomas.

Histomorphologic Features of Spontaneous and Chemically-Induced Pulmonary Neoplasms in B6C3F1 Mice and Fischer 344 Rats*1

The histomorphologic features of spontaneous and chemically-induced lung neoplasms in male and female B6C3F1 mice and Fischer 344 rats are described. Primary pulmonary neoplasms in mice and rats were classified as alveolar/bronchiolar (A/B) adenoma or carcinoma (including variants with squamous and mucinous cell differentiation), bronchial adenoma or carcinoma, squamous cell carcinoma or mesenchymal tumors. A/B adenomas and carcinomas were the most common spontaneous pulmonary neoplasms observed in both mice and rats, but were observed less frequently in rats. In the National Toxicology Program (NTP) historical control database the incidence of spontaneous A/B adenomas in male (n = 2,084) and female (n = 2,079) mice is 13.8% and 4.9%, respectively; for A/B carcinomas, it is 5.3% and 2.4%, respectively. In male (n = 3,877) and female (n = 3,919) rats, spontaneous pulmonary neoplasms are rare with historical control rates less than 3% for A/B adenomas or carcinomas in either sex. The spontaneous A/B adenomas and carcinomas observed in mice and rats typically had papillary, solid or mixed (papillary and solid) histologic growth patterns. Pulmonary neoplasms from mice and rats treated with chemical carcinogens reviewed from 2-year studies consisted primarily of A/B adenomas and carcinomas. These tumors had papillary, glandular/tubular, solid or mixed (combination of 2 or more) histologic growth patterns. A few of the A/B neoplasms had areas of squamous or mucinous cell differentiation. Other less frequently occurring spontaneous and chemically-induced neoplasms included squamous cell carcinomas, bronchial adenomas and carcinomas, and sarcomas.

Increased incidence of spontaneous and 2-acetylaminofluorene-induced liver and bladder tumors in B6C3F1 mice fed AIN-76A diet versus NIH-07 diet

Diet is a major influence on the responses of experimental animals to drugs, toxins, and carcinogens. Two diets used widely in toxicological and carcinogenic studies, and considered to be nutritionally adequate, were compared with respect to neoplastic responses to 2-acetylaminofluorene (2-AAF). Both sexes of weanling B6C3F1 mice were fed either AIN-76A (a purified diet) or NIH-07 (a natural ingredient diet), with or without 2-AAF, for up to 2 years. Dosages of 2-AAF were administered to males at 0, 40, 60, or 80 ppm in each diet and to females at 0, 150, 200, or 250 ppm. Each group consisted of 96 mice, except the groups of females dosed at 0 and 150 ppm, which consisted of 120 and 72 mice, respectively. The incidence of malignant liver tumors was significantly greater in all AIN-fed groups compared to corresponding NIH-fed groups, as was the total incidence of tumors (malignant + benign). Similarly, the incidence of malignant and total bladder tumors was greater in AIN-fed groups of mice administered the two high doses of 2-AAF for each sex compared to NIH-fed groups. No malignant bladder tumors were observed among any of the groups of mice receiving control diets. This was also true for the males on low dosages (40 ppm) of 2-AAF. The AIN-fed group of low dose females (150 ppm) developed 4% incidence of malignant bladder tumors. This study dramatically showed the importance of diet selection in chronic carcinogenic studies and suggests that results obtained with the use of a purified diet may differ both qualitatively and quantitatively from results obtained with a natural ingredient diet.

Activation of Protooncogenes in Mouse Lung Tumors

The detection of activated protooncogenes in mouse lung tumors has led to a major advance in our understanding of carcinogenesis of the lung at the molecular level. A high frequency of activated K-ras protooncogenes has been detected in tetranitromethane (TNM)- and 1,3-butadiene-induced lung tumors in B6C3F1 mice. In the past several years, we have pursued protooncogene activation in spontaneous and chemically induced tumors of strain A mice. The strain A mouse has a high incidence of spontaneous lung tumors and is susceptible to tumor induction by chemical carcinogens. We have detected and characterized the activated protooncogenes in the DNA of both spontaneously occurring and chemically induced lung tumors of strain A mice. Activated K-ras genes were detected using the NIH/3T3 transfection assay, and the activating mutations were identified by utilizing the polymerase chain reaction (PCR) and direct sequence analysis. A strong selectivity of mutations in the K-ras genes were observed in chemically induced lung tumors, as compared to spontaneous tumors, indicating that the carcinogens directly induced point mutations in the K-ras protooncogene. These findings suggest that the strain A mouse lung tumor model appears to be a very sensitive system to identify the mechanism by which chemical carcinogens activate the K-ras gene in lung tissue in vivo.

Increased Incidence of Spontaneous and 2-Acetylaminofluorene-Induced Liver and Bladder Tumors in B6C3F1 Mice Fed AIN-76A Diet versus NIH-07 Diet

Increased Incidence of Spontaneous and 2-Acetylaminofluorene-Induced Liver and Bladder Tumors in B6C3F1 Mice Fed AIN-76A Diet versus NIH-07 Diet

Increased expression of c-myc and c-H-ras in dichloroacetate and trichloroacetate-induced liver tumors in B6C3F1 mice

The expression of c-myc and c-H-ras in hyperplastic nodules and hepatocellular carcinomas induced in male B6C3F1 mice after chronic administration of dichloroacetate (DCA) and trichloroacetate (TCA) was studied using in situ hybridization. Expression of c-myc and c-H-ras mRNA was increased in both nodules and carcinomas relative to surrounding tissue and tissues obtained from control animals. Myc expression was similar in hyperplastic nodules and carcinomas induced by DCA, but was significantly higher in TCA-induced carcinomas than in hyperplastic nodules and carcinomas produced by DCA. In carcinomas from animals whose TCA treatment was suspended at 37 weeks, c-myc expression remained high relative to control and surrounding liver tissue at 52 weeks. In contrast, the expression of c-H-ras was consistently elevated in carcinomas from both treatments relative to hyperplastic nodules and non-tumor tissue. Within carcinomas from both treatments, focal areas could be located which expressed even higher levels of c-myc. This heterogeneity was not observed in carcinomas hybridized to c-H-ras-probes. These data suggest that elevated expression of c-H-ras and c-myc might play an important role in the development of hepatic tumors in B6C3F1 mice. Elevated expression of c-H-ras was closely associated with malignancy. Increased c-myc expression does not seem necessary for progression to the malignant state. On the other hand, the increased expression of c-myc appears related to the earlier progression of TCA-induced tumors to the malignant state.

Liver tumor induction in B6C3F1 mice by dichloroacetate and trichloroacetate

Male and female B6C3F1 mice were administered dichloroacetate (DCA) and trichloroacetate (TCA) in their drinking water at concentrations of 1 or 2 g/l for up to 52 weeks. Both compounds induced hepatoproliferative lesions (HPL) in male mice, including hepatocellular nodules, adenomas and hepatocellular carcinomas within 12 months. The induction of HPL by TCA was linear with dose. In contrast, the response to DCA increased sharply with the increase in concentration from 1 to 2 g/l. Suspension of DCA treatment at 37 weeks resulted in the same number of HPL at 52 weeks that would have been predicted on the basis of the total dose administered. However, none of the lesions in this treatment group progressed to hepatocellular carcinomas. Conversely, the yield of HPL at 52 weeks when TCA treatment was syspended at 37 weeks was significantly below that which would have been predicted by the total dose administered. In this case, 3 of 5 remaining lesions were hepatocellular carcinomas. Throughout active treatment DCA-treated mice displayed greatly enlarged livers characterized by a marked cytomegaly and massive accumulations of glycogen in hepatocytes throughout the liver. Areas of focal necrosis were seen throughout the liver. TCA produced small increases in cell size and much a more modest accumulation of glycogen. Focal necrotic damage did not occur in TCA-treated animals. TCA produced marked accumulations of lipofuscin in the liver. Lipofuscin accumulation was less marked with DCA. These data confirm earlier observations that DCA and TCA are capable of inducing hepatic tumors in B6C3F1 mice and argue that the mechanisms involved in tumor induction differ substantially between these two similar compounds. Tumorigenesis by DCA may depend largely on stimulation of cell division secondary to hepatotoxic damage. On the other hand, TCA appears to increase lipid peroxidation, suggesting that production of radicals may be responsible for its effects.

Recent Scientific Developments That Affect the Assessment of Risk Posed by Trichloroethylene and Perchloroethylene

Abstract Trichloroethylene (TRI) and perchloroethylene (PER) are chlorinated hydrocarbons with similar chemical and toxicological properties. Cancer bioassay results with TRI and PER have shown increased incidence of liver tumors in B6C3F1 mice. Though the exact mechanism of tumor induction caused by TRI or PER is unknown, metabolic activation is strongly implicated. The major metabolite, trichloroacetic acid (TCA), induces peroxisomal proliferation (PP) in livers of rodents. Evidence suggests that PP in the liver is linked to the development of hepatic carcinomas. Whether PP is causative or just a by-product of liver cancer is unknown, but results establish that human cells do not react to TCA as rodent cells do. The current information on pharmacokinetics and mechanism-of-action for TRI and PER do not support a concern for carcinogenicity of exposed persons, based on findings of cancer in rodents. Moreover, the findings do not suggest a need for quantitative risk assessment.

SPONTANEOUS THYROID, PARATHYROID, AND PANCREATIC ISLET TUMORS IN F344 RATS AND B6C3F<sub>1</sub> MICE

The incidence and histopathological features of spontaneous thyroid, parathyroid, and pancreatic islet tumors encountered in long-term oncogenicity studies using F344 rats or B6C3F1 mice were investigated. Spontaneous C-cell tumors of the thyroid were found at high incidence in both sexes of F344 rats, whereas pancreatic islet-cell tumors were only observed at high incidence in males. All tumors were usually found in old animals, and were rare in young and middle aged rats. In general, the incidences of endocrine tumors in B6C3F1 mice were very much lower, although follicular cell tumors of the thyroid were sometimes observed. In the endocrine system, the follicular cell of the thyroid is the most susceptible to chemical carcinogens. Carcinogen-induced tumors can be distinguished from spontaneous lesions due to their higher incidence, earlier induction, multiplicity, and increased evidence of invasion and metastasis.

Ovarian Neoplasms in F344 Rats and B6C3F1 Mice

The National Toxicology Program (NTP) classification system for rat and mouse ovarian tumors is presented. The classification system is based on previous classification systems and on a review of all the primary ovarian tumors from the archives of the National Cancer Institute (NCI) and NTP Carcinogenesis Testing Programs. The relative frequency and principal diagnostic features of 204 ovarian tumors from 39,851 female F344 rats and 587 ovarian tumors from 41,102 female B6C3F1 mice are described. The most frequently observed neoplasms in F344 rats were malignant granulosa cell tumors (29% of primary rat ovarian neoplasms observed), benign undifferentiated sex cord-stromal tumors (26%), benign granulosa cell tumors (16%), and benign Sertoli cell tumors (7%). The most frequent neoplasms in B6C3F1 mice were cystadenomas (24%), tubulostromal adenomas (24%), benign granulosa cell tumors (21%), and benign teratomas (8%).

Ovarian neoplasms in F344 rats and B6C3F1 mice.

The National Toxicology Program (NTP) classification system for rat and mouse ovarian tumors is presented. The classification system is based on previous classification systems and on a review of all the primary ovarian tumors from the archives of the National Cancer Institute (NCI) and NTP Carcinogenesis Testing Programs. The relative frequency and principal diagnostic features of 204 ovarian tumors from 39,851 female F344 rats and 587 ovarian tumors from 41,102 female B6C3F1 mice are described. The most frequently observed neoplasms in F344 rats were malignant granulosa cell tumors (29% of primary rat ovarian neoplasms observed), benign undifferentiated sex cord-stromal tumors (26%), benign granulosa cell tumors (16%), and benign Sertoli cell tumors (7%). The most frequent neoplasms in B6C3F1 mice were cystadenomas (24%), tubulostromal adenomas (24%), benign granulosa cell tumors (21%), and benign teratomas (8%).

Morphology and Classification of Ovarian Neoplasms in F344 Rats and (C57BL/6 × C3H)F<sub>1</sub> Mice

For the updating of the National Toxicology Program (NTP) classification system for rat and mouse ovarian tumors, all the primary ovarian tumors from the archives of the National Cancer Institute and NTP Carcinogenesis Testing Programs were reviewed. The relative frequency and principal diagnostic features of 204 ovarian tumors from 39,851 female F344 rats and of 587 ovarian tumors from 41,102 female (C57BL/6 X C3H)F1 (B6C3F1) mice were described. The most frequently observed neoplasms in F344 rats were malignant granulosa cell tumors (29% of primary rat ovarian neoplasms observed), benign undifferentiated sex cord-stromal tumors (26%), benign granulosa cell tumors (16%), and benign Sertoli cell tumors (7%). The most frequent neoplasms in B6C3F1 mice were cystadenomas (24%), tubulostromal adenomas (24%), benign granulosa cell tumors (21%), and benign teratomas (8%). Ovarian neoplasms were not significantly related to treatment in rats. Tubulostromal adenomas, benign and malignant granulosa cell tumors, and benign teratomas were significantly more frequent in treated mice than in controls.

Ovarian choriocarcinoma in the mouse.

Choriocarcinoma is one of the rarest ovarian tumors in any animal species. This paper describes the gross and microscopic appearance of seven such neoplasms in B6C3F1 mice. Mean age at death was 47 weeks. Tumors were described at necropsy as dark or hemorrhagic cystic lesions. On microscopic examination tumors were composed of hematocysts, intercellular hemorrhage, and cytotrophoblasts, syncytiotrophoblasts, and/or trophoblastic giant cells. Cytotrophoblasts, syncytiotrophoblasts, and occasional giant cells were present in three cases while the other four tumors contained only trophoblastic giant cells.

Liver lesions in B6C3F1 mice: the National Toxicology Program, experience and position.

The spectrum of hepatocellular proliferative lesions in B6C3F1 (C57BL/6N X C3H/HeN MTV-) mice used in National Toxicology Program (NTP) two-year carcinogenicity studies includes foci of cellular alteration, adenoma, carcinoma, and hepatoblastoma. The diagnosis of hepatocellular hyperplasia is reserved for those non-neoplastic proliferative lesions which are believed to occur secondary to necrosis or a degenerative process in the liver. Diagnostic criteria, similar to those previously published, are used during the NTP peer review of these lesions. Incidences of liver tumors in control mice have been established from the NTP database and the frequency of enhanced liver tumor responses in treated mice has been determined based on 278 two-year carcinogenicity studies. Although alternative rodent strains are being examined, at the present time there is no acceptable alternative to the B6C3F1 mouse for NTP carcinogenicity studies. Interim sacrifices and "stop" studies are incorporated into two-year studies to gain knowledge about the biological behavior of mouse liver neoplasia. Also, data are being gathered relative to the pattern of oncogene activation in spontaneous and chemically induced liver tumors in B6C3F1 mice.

DNA-binding assay of methyl chloride.

Fischer-344 rats and B6C3F1 mice of both sexes were exposed in closed chambers to 14C-labeled methyl chloride. Different clearance values from the gas phase of the system indicated that, based on body weight, mice metabolized the test compound much faster than rats. After isolation of DNA and nucleoproteins from liver and kidneys radioactivity was found in all macromolecular samples; this was ascribed to metabolic C1-incorporation. Radioactivity incorporation was particularly high in DNA of mouse kidneys, suggesting a high turnover to active C1 bodies (formaldehyde, formate) in this tissue. Analyses of DNA samples from kidneys of female and male mice showed neither 7-N-methylguanine nor O6-methylguanine. Hence, the formation of tumors in B6C3F1 mice exposed to high concentrations of methyl chloride is not based on methylation of DNA in this tissue.

Morphology and sex-dependent manifestation of nasal tumors in B6C3F1 mice after chronic inhalation of 1,2-dibromo-3-chloropropane.

Groups of 50 B6C3F1 mice of each sex were exposed to air containing of 1,2-dibromo-3-chloropropane at concentrations of 0.6 or 3.0 ppm for 6 hours per day, 5 days per week for 103 weeks. Untreated controls consisted of 50 mice of each sex. All surviving animals were killed at 104 weeks. Adenomas, squamous cell papillomas, adenocarcinomas, squamous cell carcinomas, carcinosarcomas, fibrosarcomas, and hemangiosarcomas of the nasal cavity occurred in male and female mice with dose related trends. Eighty-seven percent of the high dose female mice had nearly twice as many tumors as the high dose males (47%). Fourtynine percent of malignant neoplasms of the high dose female mice infiltrated the brain or metastasized to the lungs. Whereas two animals (4%) of the low dose female mice showed infiltrating carcinomas, no male mice exhibited this feature.