title: Carcinogenesis bioassay of trichloroethylene.
abstract: Trichloroethylene (TCE), a halogenated chemical, has been tested for carcinogenicity in the National Cancer Institute's Carcinogenesis Bioassay Program. Trichloroethylene has been used primarily as a solvent in industrial degreasing operations. Other uses have been as a solvent in dry cleaning and food processing, as an ingredient in printing inks, paints. etc., and as a general anesthetic or analgesic. Industrial grade (>99% pure) trichloroethylene was tested using 50 animals per group at 2 doses and with both sexes of Osborne-Mendel rats and B6C3F1 mice. Twenty of each sex and species were maintained as matched controls, in addition to colony and positive carcinogen controls. Animals were exposed to the compound by oral gavage 5 times per week for 78 weeks. At the end of treatment, animals were observed until terminal sacrifice at 110 weeks for rats and 90 weeks for mice. A complete necropsy and microscopic evaluation of all animals (except 7 of the original 480) was conducted. Two doses were used with animals started on test at approximately 6 weeks of age. The initial doses used in this test were the estimated maximum tolerated dose (MTD) and 1/2 MTD, as predicted from data obtained in a 6-week toxicity study. For rats, the initial doses were 1,300 and 650 mg/kg body weight. These were changed, based upon survival and body weight data, so that the "time-weighted average" doses were 549 and 1,097 mg/kg for both male and female rats. For mice, the initial doses were 1,000 and 2,000 mg/kg for males and 700 and 1,400 mg/kg for females. The doses were increased so that the "time-weighted average" doses were 1,169 and 2,339 mg/kg for male mice and 869 and 1,739 mg/kg for female mice. Clinical signs of toxicity, including reduction in weight, were evident in treated rats. These, along with an increased mortality rate necessitated a reduction in doses during the test. In contrast, very little evidence of toxicity was seen in mice, so doses were increased slightly during the study. The increased mortality in treated male mice appears related to the presence of liver tumors. A variety of neoplastic lesions were observed in rats with no significant difference between trichloroethylene-treated and control animals. The only lesion that might be attributed to the treatment was a chronic nephropathy found in both sexes and at both dose levels. With both male and female mice, primary malignant tumors of the liver, i.e., hepatocellular carcinoma, were observed in high numbers. For males, 26/50 low dose and 31/48 high dose animals had hepatocellular carcinomas as compared with 1/20 matched controls and 5/77 colony controls. The differences between treated and matched control males at both doses were highly significant (P<0.01). For females, hepatocellular carcinomas were observed in 4/50 low dose and 11/47 high dose animals as compared with 0/20 matched controls and 1/80 colony controls. While the difference between the high dose female mice and matched controls was also highly significant (P<0.01), the difference at the low dose was less (P=0.09). For both male and female mice, age-adjusted tests for linear trend (dose response) were highly significant for hepatocellular carcinoma (P<0.001 for males and P=0.002 for females). In male mice at the high doses, hepatocellular carcinomas were observed early in the study. The first was seen at 27 weeks; 9 others were found in male mice dying by the 78th week. The tumor was not observed so early in low dose male or female mice. The diagnosis of hepatocellular carcinoma was based on size, histologic appearance, and presence of metastasis, especially to the lung. No other lesion was significantly elevated (P<0.05) in treated mice. The incidence of hepatocellular carcinomas in the trichloroethylene-matched controls was typical of that observed in colony controls. Carbon tetrachloride (CCl4) was used as a positive control for the series of chlorinated chemicals which included trichloroethylene. While virtually all male and female mice developed hepatocellular carcinomas following carbon tetrachloride treatment, the response in the Osborne-Mendel rats was considerably less. Only about 5% developed hepatocellular carcinomas. Thus, there appears to be a marked difference in sensitivity to induction of carcinomas by chlorinated compounds between the B6C3F1 mouse and the Osborne-Mendel rat. The results of this carcinogenesis test of trichloroethylene clearly indicate that trichloroethylene induced a hepatocellular carcinoma response in mice. While the absence of a similar effect in rats appears most likely attributable to a difference in sensitivity between the Osborne-Mendel rat and the B6C3F1 mouse, the early mortality of rats due to toxicity must also be considered.