Clear Evidence Of Carcinogenicity Research Articles

A weight of evidence evaluation of the mode of action of isoeugenol

Isoeugenol is one of several phenylpropenoid compounds that is used as a fragrance, food flavoring agent and in aquaculture as a fish anesthetic. Carcinogenicity testing in rats and mice by NTP resulted in clear evidence of carcinogenicity (hepatic adenomas/carcinomas) in male mice only. A nongenotoxic threshold mode of action (MOA) is postulated for isoeugenol and is discussed considering the IPCS MOA and Human Relevance Framework. The weight of evidence indicates that isoeugenol is not genotoxic and that the carcinogenic outcome in male mice relates directly to the metabolism of individual compounds. Benchmark Dose (BMD) modeling was conducted to determine a Point of Departure (POD) and potential threshold of carcinogenicity. The results of the BMD evaluation for isoeugenol resulted in an estimated POD for carcinogenicity in the male mouse of 8 mg/kg with a lower limit of 4 mg/kg, representing a POD for the determination of an acceptable daily intake. With application of uncertainty factors, an ADI of 40 μg/kg is calculated. This daily dose in humans would be protective of human health, including carcinogenicity. A corresponding maximum residual level (MRL) of 3200 μg/kg fish is also estimated based on this POD that considers the threshold MOA.

Chemical-Induced Oral Squamous Cell Neoplasms in Rodents: An Overview of NTP 2-Year Cancer Studies.

Oral cancer is the seventh most common malignancy worldwide, and lifestyle factors participate in its development. Rodent studies can help identify substances that contribute to its development and provide information on the early stages of carcinogenicity. The National Toxicology Program (NTP) has conducted more than 500 short-term and 2-year toxicology and carcinogenicity studies in rodents, and some of the tested compounds resulted in oral cancer. Our goal was to review the NTP carcinogenic studies to describe those chemicals that have oral carcinogenic outcome in rodents. For this project, we reviewed the results from all NTP carcinogenicity studies and a board-certified veterinary pathologist reviewed the slides from all neoplasms in the oral cavity that were considered treatment related. We have identified 26 chemicals with an adverse effect in the oral cavity. Fourteen chemicals demonstrated clear evidence of carcinogenicity in the oral cavity. We provide information on the carcinogenic findings in rodents together with a detailed description of the morphologic aspects of the oral cancers and speculate that the carcinogenic effects can be induced by different pathological modes of action. The findings reviewed here provide indicators for potential oral carcinogenesis processes in rodent models, which can be further investigated in future mechanistic studies.

Biocompatibility and Carcinogenicity of Carbon Nanotubes as Biomaterials.

With the development of nanotechnology in recent years, there have been concerns about the health effects of nanoparticles. Carbon nanotubes (CNTs) are fibrous nanoparticles with a micro-sized length and nano-sized diameter, which exhibit excellent physical properties and are widely studied for their potential application in medicine. However, asbestos has been historically shown to cause pleural malignant mesothelioma and lung cancer by inhalation exposure. Because carbon nanotubes are also fibrous nanotubes, some have raised concerns about its possible carcinogenicity. We have reported that there is no clear evidence of carcinogenicity by local and intravenous administration of multi-walled CNTs to cancer mice models. We firmly believe that CNTs can be a safe, new, and high-performance biomaterials by controlling its type, site of administration, and dosage.

Long-term toxicology and carcinogenicity of 2,4,6-trichlorophenol

Carcinogenesis bioassays were conducted by giving 2,4,6-trichlorophenol [2,4,6-TCP] in feed to groups of 50 male and female Fischer rats and male B6C3F1 mice for two years. Dietary concentrations were 0 [20/group], 5000 [0.5%], or 10,000 [1%] ppm. Female mice began with 10,000 and 20,000ppm but after 38weeks were lowered due to reduced body weights to 2500 and 5000ppm for 67weeks; exposures averaged 5200 and 10,400ppm. Adverse effects at two years were leukocytosis and monocytosis of peripheral blood and hyperplasia of bone marrow in both sexes of rats. In mice, liver toxicity, including individual liver cell abnormalities, focal areas of cellular alteration, and focal and nodular areas of hyperplasia were commonly present. Regarding carcinogenic activity, TCP caused leukemias/lymphomas in male rats, and possibly in female rats and female mice as well, and induced liver tumors in male and female mice. Using NTP categories of evidence indicates ‘clear evidence of carcinogenicity’ for male rats [hematopoietic system tumors]; ‘equivocal evidence of carcinogenicity’ for female rats [hematopoietic system tumors]; ‘clear evidence of carcinogenicity’ for male and female mice [liver tumors].

Carcinogenesis studies of cresols in rats and mice

Cresols, monomethyl derivatives of phenol, are high production chemicals with potential for human exposure. The three isomeric forms of cresol are used individually or in mixtures as disinfectants, preservatives, and solvents or as intermediates in the production of antioxidants, fragrances, herbicides, insecticides, dyes, and explosives. Carcinogenesis studies were conducted in groups of 50 male F344/N rats and 50 female B6C3F1 mice exposed to a 60:40 mixture of m- and p-cresols ( m-/ p-cresol) in feed. Rats and mice were fed diets containing 0, 1500, 5000, or 15,000 ppm and 0, 1000, 3000, or 10,000 ppm, respectively. Survival of each exposed group was similar to that of their respective control group. Mean body weight gains were depressed in rats exposed to 15,000 ppm and in mice exposed to 3000 ppm and higher. A decrease of 25% over that of controls for the final mean body weight in mice exposed to 10,000 ppm appeared to be associated with lack of palatability of the feed. A marginally increased incidence of renal tubule adenoma was observed in the 15,000-ppm-exposed rats. The increased incidence was not statistically significant, but did exceed the range of historical controls. No increased incidence of hyperplasia of the renal tubules was observed; however, a significantly increased incidence of hyperplasia of the transitional epithelium associated with an increased incidence of nephropathy was observed at the high exposure concentration. The only significantly increased incidence of a neoplastic lesion related to cresol exposure observed in these studies was that of squamous cell papilloma in the forestomach of 10,000-ppm-exposed mice. A definitive association with irritation at the site-of-contact could not be made because of limited evidence of injury to the gastric mucosa at the time of necropsy. However, given the minimal chemical-related neoplastic response in these studies, it was concluded that there was no clear evidence of carcinogenicity in male rats or female mice exposed to the cresol mixture.

Inhalation Carcinogenicity and Chronic Toxicity of Carbon Tetrachloride in Rats and Mice

Carcinogenicity and chronic toxicity of carbon tetrachloride were examined by inhalation exposure of 50 F344 rats and 50 BDF1 mice of both sexes to carbon tetrachloride at 0 (clean air), 5, 25, or 125 ppm (v/v) for 6 h/day, 5 days/wk, for 104 wk. Incidences of hepatocellular adenomas and carcinomas in rats and mice of both sexes and of adrenal pheochromocytomas in mice of both sexes were significantly increased dose-dependently. Hepatocellular carcinomas and cirrhosis significantly occurred in the 125-ppm-exposed rats of both sexes, and 3 cases of hepatocellular carcinomas and increased incidences of hepatic altered cell foci were noted in the 25-ppm-exposed female rats. Hepatocellular carcinomas were induced in mice of both sexes at 25 and 125 ppm, and hepatocellular adenomas occurred in females at 5 ppm without any degenerative or necrotic change in hepatocytes. Hepatocellular carcinomas metastasized to the lung. The chronic hepatotoxicity was characterized by cirrhosis, fibrosis, and fatty change in rats, and ceroid deposition, bile-duct proliferation, and hydropic change in mice. Survival rates were decreased in the 125-ppm-exposed rats and mice of both sexes and in the 25-ppm-exposed female mice, in association with decreased body weights. The decreased survival rates were considered to be causally related to both various tumors including hepatocellular carcinomas and severe chronic progressive nephropathy in rats and to hepatocellular carcinomas in mice. This study provided clear evidence of carcinogenicity for carbon tetrachloride in rats and mice. A cytotoxic-proliferative and genotoxic mode of action for carbon tetrachloride-induced hepatocarcinogenesis was suggested.

Are tumor incidence rates from chronic bioassays telling us what we need to know about carcinogens?

Chronic bioassays for over 500 chemicals have been conducted under the auspices of the National Cancer Institute and/or the National Toxicology Program (NTP) to screen chemicals for carcinogenicity, providing a wealth of information about bioassays. Typically, chemicals are administered for two years to both sexes in each of one strain of rats and mice generally at the maximum tolerated dose (MTD), MTD/2, MTD/4 (in recent years), as well as unexposed control animals. In an attempt to ascertain the sensitivity of this bioassay to detect animal carcinogens tested at the MTD for the current experimental design, the false negative rate (failure to detect increased tumor rates) was investigated. This was accomplished by examining the tumor incidences from over 150 NTP bioassays and estimating the probability that a statistically significant ( P ⩽ 0.01) dose–response trend would be obtained at one or more tissue sites in either sex of rats or mice if 200, rather than 50, animals were used per dose group. This provides an estimate of the proportion of chemicals that were not declared high-dose animal carcinogens due to the limited sample size of 50 animals per species–sex–dose group. In this series of chemicals tested, 97/156 (62%) were identified by the NTP to show some or clear evidence of carcinogenicity. With an increase of the number of animals per dose group from 50 to 200, it is estimated that 92% of these chemicals would show statistically significant ( P ⩽ 0.01) dose–response trends at one or more tissue sites in either sex of rats or mice. Many of these chemicals are not genotoxic. Some chemicals had no structural alerts for carcinogenicity, but were tested because of potentially high human exposure. This analysis suggests that almost all of the chemicals selected would produce a statistically significant increase in tumor incidence at the MTD with larger sample sizes. Hence, this MTD bioassay screen is not distinguishing between true carcinogens and non-carcinogens. Rather, the screen is simply failing to detect the weaker carcinogens at the MTD. More than 30% of chemicals tested failed to detect statistically significant dose–response trends for tumors because of inadequate sample sizes of 50 animals per dose. Presumably, little or no action would have been taken to regulate exposures to these chemicals as potential carcinogens due to lack of a positive test result. This analysis does not suggest that most chemicals are carcinogenic at human exposure levels nor does it suggest that more than 50 animals should be tested per dose group. With an MTD that may produce a difference (up to 10%) in weight gain between treated and control animals, there quite possibly is cytotoxicity at the MTD. Increased carcinogenicity would be expected from increased opportunities for mutagenic activity during regenerative cell replication to compensate for cytotoxicity. Since it appears that almost all chemicals tested adequately at the MTD will demonstrate carcinogenicity, it is tempting to surmise that this is due in large part to one or more nearly universal modes of action, such as, regenerative cell replication at the MTD rather than due to some unique carcinogenic property of a chemical. That is, the current bioassay possibly is just primarily a screen for the more potent cytotoxins at the MTD, rather than a screen specifically for carcinogenicity. This issue should be examined and suggests that cytotoxicity and cell proliferation should be considered in setting the MTD, particularly for non-genotoxic (non-DNA reactive) chemicals. From a public health view, it is prudent to assume that most chemicals could demonstrate increased tumor incidence rates at the MTD in rodents. The current standard NTP bioassay provides sufficient data to estimate a benchmark dose associated with a specified low tumor incidence to be used as a point-of-departure for cancer risk assessments. The question that should be investigated by a bioassay is not whether a chemical is a carcinogen at the MTD, but what is the relationship between dose and cytotoxicity and/or other modes of action that could produce an excess of tumors?

Genotoxicity studies with chloroethane

In a recent 2-year inhalation study with F344 rats and B6C3F1 mice conducted as part of the U.S. National Toxicology Program (NTP, 1989), chloroethane (EC1) at an exposure concentration of 15 000 ppm induced a high incidence of endometrial uterine carcinomas only in female mice but not in rats, leading to the conclusion of “clear evidence of carcinogenicity” for the mouse. In order to elucidate whether a genotoxic effect may be a critical factor for the carcinogenicity of EC1 in the mouse, we have performed three genotoxicity tests: (1) in vitro HPRT test with CHO cells according to a specially developed gas protocol, (2) in vivo/in vitro UDS with female B6C3F1 mice at an exposure concentration of 25 000 ppm (6 h/day, 3 days), (3) in vivo micronucleus assay with male and female B6C3F1 mice exposed to 25 000 ppm EC1 according to the same schedule. In the in vitro HPRT test a mutagenic potential of EC1 was detected in the presence as well as in the absence of S9 mix. In contrast, both in vivo test systems failed to detect any indications of genotoxicity of chloroethane at an exposure concentration even higher than that of the NTP study. It is suggested that in vivo the genotoxic potential of EC1 is so low that an assumed genotoxic damage is below the detection limit of the test systems used. This leads to the conclusion that genotoxicity may not be a key factor in the induction of the uterine carcinomas in the B6C3F1 mouse.

Nasal Cavity Neoplasia in F344/N Rats and (C57BL/6 × C3H)F<sub>1</sub> Mice Inhaling Propylene Oxide for up to Two Years<xref ref-type="fn" rid="FN1">2</xref>

Propylene oxide (CAS: 75-56-9) was studied for potential carcinogenicity and chronic toxicity by inhalation in F344/N rats and (C57BL/6 x C3H)F1 mice. Groups of 50 animals of each sex were exposed to 0, 200, or 400 ppm propylene oxide for 6 hours/day, 5 days/week, for up to 103 weeks. Survival decreased in mice exposed to propylene oxide; the decrease was significant (P less than .005) in mice exposed to 400 ppm. Survival of exposed rats was comparable to that of controls. Mean body weight of rats and mice exposed to 400 ppm propylene oxide decreased, when compared to that of controls, during the 2d year of exposure. Exposure to propylene oxide for up to 2 years induced inflammatory and proliferative responses in nasal cavity of both species. There was clear evidence of carcinogenicity in mice exposed to 400 ppm propylene oxide; 10 of 50 males and 5 of 50 females had hemangiomas or hemangiosarcomas of the nasal submucosa. Papillary adenomas involving the nasal respiratory epithelium and underlying submucosal glands were observed in 3 female rats and 2 male rats exposed to 400 ppm propylene oxide.