Liver Tumors In Animals Research Articles

Mouse models of liver cancer: Progress and recommendations.

To clarify the pathogenesis of hepatocellular carcinoma (HCC) and investigate the effects of potential therapies, a number of mouse models have been developed. Subcutaneous xenograft models are widely used in the past decades. Yet, with the advent of in vivo imaging technology, investigators are more and more concerned with the orthotopic models nowadays. Genetically engineered mouse models (GEM) have greatly facilitated studies of gene function in HCC development. Recently, GEM of miR-122 and miR-221 provided new approaches for better understanding of the in vivo functions of microRNA in hepatocarcinogenesis. Chemically induced liver tumors in animals share many of the morphological, histogenic, and biochemical features of human HCC. Yet, the complicated and obscure genomic alternation restricts their applications. In this review, we highlight both the frequently used mouse models and some emerging ones with emphasis on their merits or defects, and give advises for investigators to chose a "best-fit" animal model in HCC research.

Combined GM-CSF and IL-12 gene therapy synergistically suppresses the growth of orthotopic liver tumors

Immunotherapy is often effective only for small tumor burdens and, in many cases, is restricted to subcutaneous tumors. Here, we investigated the antitumor effects of combination therapy with GM-CSF and IL-12 on orthotopic liver tumors with intermediate or large tumor volumes, or on chemically-induced multifocal liver tumors in animals. Adenoviruses encoding GM-CSF or IL-12 were injected intratumorally to animals bearing transplanted tumors, or injected via intrahepatic artery in animals with primary multifocal liver tumors induced by diethylnitrosamine. Our results demonstrated that IL-12, but not GM-CSF, monotherapy displayed significant therapeutic effects, whereas combination therapy with both cytokines displayed synergistic antitumor effects not only on transplanted tumor models with intermediate or large tumor loads, but also on carcinogen-induced multifocal liver tumors. Effector cell analyses, revealed by in vivo cell subset depletion, flow cytometry analysis, and immunohistochemical staining of tumor infiltrates, indicated that NK cells were the prominent antitumor effectors for the IL-12-mediated antitumor activity, whereas CD8+ T cells, NKT cells, and macrophages were more important than NK cells in the combination therapy-mediated antitumor effects. Both IL-12 monotherapy and combination therapy could induce various types of effectors and high levels of IFN-gamma; however, the latter induced much higher levels than the former, which may explain why combination therapy is superior to IL-12 monotherapy. Combination therapy with GM-CSF and IL-12 represents a promising immunotherapy strategy for treating orthotopic, widespread liver tumors.

Roles of reactive oxygen species in hepatocarcinogenesis and drug resistance gene expression in liver cancers

Hepatocellular carcinoma (HCC) has traditionally been an attractive system for cancer research because many animal HCC models are available. It is well known that liver tumors in animals can be induced by many different protocols, such as chronic hepatitis viral infections, carcinogens, toxins, steroid hormones, and dietary intervention. Although these different inducers have different cellular targets and modes of cytotoxic effects, their common denominator is the formation of reactive oxygen species (ROS). In this review, we present compelling evidence to support the hypothesis that ROS play important roles in hepatocarcinogenesis and the associated upregulation of drug resistance gene expression.

EVALUATION OF THE HEALTH RISK ASSOCIATED WITH EXPOSURE TO CHLOROFORM IN INDOOR SWIMMING POOLS

The exposure of swimmers to chloroform (CHCl5) was investigated in indoor swimming pools of the Quebec City region along with the associated carcinogenic risk. Six training sessions involving 52 competition swimmers (11 to 20 yr old) were conducted in 3 different pools, while 12 adult leisure swimmers attended 5 sessions, each held in a different pool. For each session, water and ambient air CHCl3 concentrations were measured and CHCl3 levels in alveolar air samples (CHCl3ALV) collected from swimmers prior to entering the swimming pool premises and after 15, 35, and 60 min of swimming. Mean water concentrations varied from 18 µg/L to 80 µg/L, while those in air ranged from 78 µg/m3 to 329 µg/m3. Multiple linear regression analyses revealed that CHCl3ALV values in competition swimmers were strongly correlated to ambient air and water levels, and to a lesser degree to the intensity of training. Only ambient air concentration was positively correlated to CHCl3ALV in the leisure group. Concentrations of CHCl3 metabo lites bound to hepatic and renal macromolecules, estimated using a physiologically based pharmacokinetic (PBPK) model, were 1.6 and 1.9 times higher for the competition swimmers than for the leisure swimmers, respectively. The highest hepatic concentration predicted in competition swimmers, 0.22 µg CHCl equivalents/kg of tissue, was at least 10,000 times lower than the smallest no observed3effect level for liver tumors in animals. Data indicate that the safety margin is therefore very large, for competitive swimmers as well as for leisure swimmers.

Use of Probabilistic Expert Judgment in Uncertainty Analysis of Carcinogenic Potency

A new approach to characterizing the state of knowledge about carcinogenic potency is described. In this approach, the carcinogenic risk posed by a specific dose is characterized by a probability distribution, indicating the relative likelihood of different risk estimates. The approach utilizes expert judgment and a probability tree and is illustrated in a case study of chloroform exposure. Experts in cancer biology/toxicology, pharmacokinetics, and dose-response modeling were identified by a panel of science-policy specialists. In a workshop, experts reviewed the chloroform data, received training in probability elicitation, and constructed a consensual probability tree based on biological theories of cancer causation. Distributions of carcinogenic risk were developed based on the probability tree, chloroform data, judgmental probabilities provided by the experts, and classical statistical techniques. Risk distributions varied considerably between experts, with some predicting essentially no risk from 100 ppb chloroform in drinking water while others have at least some probability on risks generally considered of regulatory significance. Estimated human risk was much lower when extrapolating from liver tumors in animals than from kidney tumors. Issues of scientific disagreement leading to different risk distributions between experts are discussed. The resulting risk distributions are compared to standard EPA risk calculations for the same exposure scenario as well as to the expert judgments of epidemiologists about cancer risks of chlorinated drinking water. Issues in combining expert judgments are discussed, and several alternative methods are presented. Strengths and weaknesses of the distributional approach are discussed.

Implications of the Carcinogenic Hazard of Low Doses of Three Hepatocarcinogenic N‐Nitrosamines1

The data of a large-scale experiment on single and combination effects of very low doses of the hepatocarcinogenic N-nitrosamines N-nitrosodiethylamine (NDEA), N-nitrosopyrrolidine (NPYR), and N-nitrosodiethanolamine (NDElA) were modeled by new statistical methods to derive implications of the carcinogenic hazard of dose ranges so low as to result in long-term toxic effects only slightly different from the background. According to this model a linear relationship was found to exist between daily exposure to low N-nitrosamine levels and time to death with liver tumor. Extrapolation of these data to zero exposure suggested the occurrence of one to ten percent of spontaneous liver tumors in animals surviving more than 1000 days. At this advanced age a further reduction of carcinogen-induced liver tumor incidence does not contribute to a longer overall survival due to competitive, probably independent, causes of death. A quasi-threshold in terms of a "no-observed-effect level" can thus be derived from the data. The observed combination effect indicates a mere additivity in liver tumor occurrence, even at very low doses which alone cause no significant carcinogenic effect during the animal's lifetime. Within the combination of NDEA, NPYR and NDElA, N-nitrosodiethylamine was found to be the most carcinogenic agent: it contributes by at least 17 orders of magnitude more to the relative risk of dying with liver tumor than the other two compounds, if the daily dose is increased by one unit (0.1 mg/kg). Likewise, NDEA shows the steepest slope when assessing the relationship between daily carcinogen doses and time to liver tumor occurrence; relative to NDElA--the least potent carcinogen on a weight basis--and NPYR a 40-fold and 9-fold quicker appearance of liver tumors has to be expected, if the daily doses are increased by an equivalent amount.

Liver Tumors in Rodents: Extrapolation to Man

Man is a poor model for the prediction of agents that are hepatocarcinogenic for laboratory rodents. Relatively few agents are known to cause any form of primary liver cancer in man. The most important is hepatitis B virus, for which there is possibly a model in the woodchuck but not one in rats or mice. The only other agents known to cause primary liver cancer in man are certain steroid hormones, vinyl chloride, and thorium dioxide. There are animal models for the first two of these and a reasonable expectation that thorium dioxide would produce liver tumors in animals if the appropriate experiments were done. Aflatoxin, a potent hepatocarcinogen in rats and other species but not mice, is strongly suspected of being an important human hepatocarcinogen in certain geographical areas of the world, but the evidence is circumstantial. There is no more than a weak association between the nutritional type of cirrhosis secondary to excessive intake of alcohol and increased primary liver cancer in man, and no evidence at all that ethanol per se causes liver tumors in mice, rats, hamsters, or mastomys. By contrast, a very large number of chemicals to which people in the West have been exposed for many decades have been found to be hepatocarcinogens in laboratory rodents. In most cases the levels of exposure required to produce liver tumors in rodents far exceed those to which man is normally exposed. The problem is to guess whether low-level exposure to such rodent hepatocarcinogens poses any real liver cancer threat to man?The mortality from primary liver cancer is very low in countries such as England and Wales where there is widespread exposure to low doses of both natural and synthetic agents which, in high dosage, cause liver tumors in rodents. This suggests that, if there is any risk, it can only be very small. Death rate data collected in England and Wales by the Registrar General are consistent with there having been a small increase in the incidence of primary liver cancer in England and Wales during the past 20 years, but the apparent increase might well be a consequence of revisions in the International Classification of Diseases system and not real. During the first half of the present century the age-standardized incidence of primary liver cancer in England and Wales was falling.(ABSTRACT TRUNCATED AT 400 WORDS)

Quantitative aspects of chemical carcinogenesis and tumor promotion in liver.

Chronic exposure of rodents to high dose levels of drugs, food additives and environmental chemicals frequently results in liver enlargement. Several of these compounds have been found to enhance the incidence of liver tumors in animals briefly exposed previously to hepatocarcinogens. Accordingly, it has been advanced that these agents act as tumor promoters. This contention has remained subject of controversy following reports that these substances may also cause liver tumors in noncarcinogen-treated rodents, particularly in those characterized by a relatively high incidence of "spontaneous" liver tumors. Since many of these chemicals are in common use, a crucial question would seem to be whether such effects are due to facilitation of the expression of pre-existing oncogenic potential, i.e., to tumor promotion, or to the synergistic action of weakly carcinogenic agents. As a result of mechanistic differences tumor promotion and syn-carcinogenesis must exhibit different dose-time-response characteristics, and, accordingly, it should be possible, in principle, to discriminate between these phenomena. However, since tumor manifestation periods in low-dose groups frequently exceed the animals average lifespan, this approach may not always yield conclusive data, unless a sensitive early marker of carcinogenic activity can be employed. There is evidence that enzyme-deficient preneoplastic areas in liver can be used for this purpose. A strong quantitative correlation between carcinogen dose, the extent of ATPase deficient areas, and the subsequent appearance of tumors has now been established for a number of hepatocarcinogens. Experimental data are consistent with the concept that two critical events (hits) are required for induction of ATPase deficiency in hepatocytes. The first hit is carcinogen-dependent, whereas the second hit would seem to be due to time-dependent event(s). Tumor-promoters, such as phenobarbital, were found to accelerate and increase formation of preneoplastic islets. This evidence, together with data indicating that the compound is devoid of carcinogenic potential, suggests that phenobarbital may be operative at relatively early stages of hepatocarcinogenesis by increasing the probability of the occurrence of the time-dependent second hit. Such effects are dose-dependent and appear to be related to the induction of liver enlargement. The changes in hepatocellular ploidy status and atypical nuclear figures observed during phenobarbital treatment and cessation thereof, suggest that this compound might induce abnormal redistributions of genetic material. It is postulated that these cytological changes may result in phenotypical manifestation of recessive oncogenic information.

Effect of various factors on the induction of liver tumors in animals by quinoline.

Tumorigenic effect of a diet containing quinoline on the liver of various animals and the synergistic or antagonistic effect by other chemicals on quinoline hepatocarcinogenesis in rats were examined. It was concluded that 4,4'-diaminodiphenylmethane (0.1%) and 3-methylcholanthrene (0.0067%) had a significant inhibitory effect on liver carcinogenesis due to quinoline in rats, but 1-naphthyl isothiocyanate (0.06%) and p-hydroxypropiophenone (1.0%) had no inhibitory effect in the present observations. Transmission electron-microscopic study demonstrated the fine structure of vascular tumors induced by quinoline. On the other hand, it was found that quinoline induced liver tumors in both sexes of mice and rats but not in hamsters or guinea pigs. Male rats were more susceptible than females to the tumorigenic action of quinoline, and mice showed the least susceptibility. Histological changes in the liver of rats or mice induced by quinoline were clasified as hemangioendotheliomas or hemangiosarcomas and hepatocellular carcinomas. Several rats treated with quinoline had hemangiosarcomatous metastatic foci in the lung.

Effect of various factors on induction of liver tumors in animals by the alpha-isomer of benzene hexachloride.

The tumorigenic effect of a diet containing the alpha-isomer of benzene hexachloride (alpha-BHC) on the liver of various animals was examined. It was found that alpha-BHC induced liver tumors in male and female mice but not in rats or hamsters in the present observations. Histological changes in the liver of mice induced by alpha-BHC were also much greater than those induced in rats or hamsters. Male animals were more susceptible to the tumorigenic action of alpha-BHC than females. Among different strains of mice, the DDY showed greatest susceptibility and the C57BL/6 showed the least. Induction oflpha-BHC was not inhibited by concomitant feeding of 1-naphthyl isothiocyanate or p- hydroxypropiophenone. However, 3-methylcholanthrene slightly inhibited their induction by alpha-BHC.