Amira Pearson

The clinical use of mutagenic anticancer drugs

Cytotoxic chemotherapy is routinely used in the treatment of cancer, and has been an important factor in increasing 5-year survival rates for some types of this disease. A range of drugs are currently available, with differing modes of action. As well as causing some direct toxic effects, most if not all of these drugs are both mutagenic and carcinogenic. Although comparative information on these properties is generally available for anticancer drugs which alkylate DNA, it has been less readily accessible for other drug classes. This special issue contains seven reviews on the mutagenic properties of the major classes of cytotoxic drugs in clinical use, as well as one on a class of drugs that is under development. Some carcinogenicity data are also summarised, where available. Additionally, there are four more general papers, including one on the use of genetic activity profiles for comparing mutagenicity of the drugs, two on germ-cell effects, and one on biomonitoring for exposure to genotoxic anticancer drugs.

Antimutagens in food plants eaten by Polynesians: micronutrients, phytochemicals and protection against bacterial mutagenicity of the heterocyclic amine 2-amino-3-methylimidazo[4,5-f]quinoline

We have previously suggested that differences in cancer incidence between Polynesians (including Maoris and people from several Pacific islands) and Europeans in New Zealand may at least partially relate to differences in the species of food plants (fruits, vegetables and cereals) preferentially eaten by these groups. Twenty-five food plants that are typically eaten in different amounts by these two population groups were selected for detailed study. Antimutagenic properties of three extracts from each of the selected plants were investigated using a preincubation mutagenicity assay with Salmonella typhimurium strain TA1538 against the mutagenicity of the heterocyclic amine 2-amino-3-methylimidazo[4,5-f]quinoline (IQ). The data revealed strong antimutagenic properties in several of the food plants commonly eaten by Polynesians, especially rice, watercress, pawpaw, taro leaves, green banana and mango. Using the New Zealand food database, a number of nutrients and micronutrients with antimutagenic and anticarcinogenic potential were identified from the selected food plants. Some of these were tested for antimutagenic potential in parallel experiments to those done with the food plant extracts. Although some of these micronutrients are antimutagens against IQ, their concentrations in the food plants failed to explain the protection against mutagenicity found in the experiments with extracts of the food plants. Thus, other types of chemical, not identified in the database, must be leading to antimutagenesis. Possible active molecules include chlorophylls, carotenoids, flavonoids and coumarins, many of which are also known to be anticarcinogens. If human cancer data are to be interpreted in terms of cancer protection, these components need urgently to be quantified in food plants in the New Zealand diet, especially in those food plants eaten in large amounts by Polynesians.

Antimutagenic effects of wheat bran diet through modification of xenobiotic metabolising enzymes

Diets containing wheat bran (WB) protect against cancers of the colon or breast in rats, and may be beneficial in humans. In a previous study of rats treated with the carcinogen 2-amino-3-methylimidazo[4,5-f]quinoline (IQ), inclusion of 10% wheat bran in the diet led to an apparent reduction in IQ metabolites but not of intact IQ in plasma. In the present study, male Wistar rats were fed diets containing 0, 10 or 20% wheat bran, and effects on xenobiotic metabolising enzymes compared. Wheat bran-supplementation showed differential effects on phase I enzymes, significantly increasing the activity of hepatic cytochrome P450 isozyme CYP3A2, but slightly reducing the activity of CYP1A1/2. The activities of both hepatic phase II detoxification enzymes glutathione-S-transferase and glucuronosyl transferase were also reduced. Western blotting revealed similar effects on expression of the proteins. Interestingly, the expression of xenobiotic metabolising enzymes (XME) in the colon appeared to be modulated independently of hepatic XME. Although the wheat bran-supplemented diet still led to an increased expression of CYP3A, it now slightly increased CYP1A in the colon. However, 20% wheat bran significantly increased the expression of both glutathione transferase isozymes, GST A1 & A2, in the colon. Natures Gold (NG) is a commercial wheat bran derivative which is lower than wheat bran in dietary fibre, but enriched in vitamins, minerals and various phytochemicals. Dietary supplementation with 20% Natures Gold led to similar trends as seen in wheat bran-fed rats, but more potent effects in both hepatic and colonic enzymes. The significance of these changes for activation of carcinogens to mutagenic metabolites was investigated using the Salmonella/mammalian microsome mutagenicity test. The activation of IQ and benzo[a]pyrene, but not cyclophosphamide, to a mutagen by hepatic S9 from wheat bran-fed or Natures Gold-fed rats was significantly reduced compared with S9 from animals on a diet lacking wheat bran. We suggest that modulation of xenobiotic metabolising enzymes may be an important component of cancer protection by wheat bran, and this effect may relate to micronutrients or cancer-protective non-nutrient phytochemicals rather more than to dietary fibre.

In vivo effects of chlorophyllin on the antitumour agent cyclophosphamide

Cyclophosphamide (CP) is a potent antitumour agent used against many forms of cancer and against certain other diseases. Chlorophyllin (CHL), which is obtained by hydrolysis of chlorophyll to remove phytyl alcohol, is an efficient antimutagenic agent and has been used as a dietary supplement or to diminish the intensity of the discomforting side effects of CP therapy. We undertook to determine the antimutagenic effectiveness of CHL against CP in a mouse model and to determine whether the antitumour efficacy of CP was compromised in vivo by CHL treatment. Experiments utilised CHL administered either in drinking water (1%) for 2 days before treatment, or by gavage (200 mg/kg) 2 hr before treatment with CP (220 mg/kg). Urinary mutagenicity following CP treatment, as determined by the Salmonella/microsome assay, was decreased by both regimes of CHL co‐treatment. Similarly, the increase in micronuclei in bone marrow polychromatic erythrocytes in response to CP was reduced by concomitant CHL treatment. In contrast, antitumour efficacy, as determined by growth delay of Colon 38 adenocarcinomas, was not diminished by CHL treatment. We conclude that CHL may have beneficial effects when used in combination with CP therapy.

Chromosomal changes in Chinese hamster AA8 cells caused by podophyllin, a common treatment for genital warts

Podophyllin, a plant derivative of variable composition, is used widely within New Zealand as a treatment for genital warts. One local source of podophyllin has been tested for ability to cause mutagenic effects in Salmonella typhimurium as well as for effects on chromosomes of Chinese hamster AA8 cells. Although only weakly mutagenic in one strain of Salmonella, podophyllin caused structural aberrations as well as changes in chromosome number in the Chinese hamster cells. The range of aberrations was similar to those caused by teniposide, a close structural relative of the major component, which was used as a positive control in the Chinese hamster cell experiments. A literature review revealed that podophyllin was shown to cause changes of chromosome number in the mouse cervix in vivo, although aberrations were not studied. Patients treated with podophyllin will usually possess one form of the papilloma virus, and this itself may have some oncogenic potential. We suggest that podophyllin could potentiate these effects and question its continued widespread use.

Photo-enhancement of the mutagenicity of 9-anilinoacridine derivatives related to the antitumour agent amsacrine

The frameshift mutagenicity of the DNA intercalating drug proflavine is known to be enhanced by photoirradiation of bacterial cultures. To determine whether this phenomenon was also present in acridine-derived antitumour drugs, cultures of Salmonella typhimurium were exposed to the antileukaemia agent amsacrine and the experimental agent N-[2-(dimethylamino)ethyl]acridine-4-carboxamide dihydrochloride (acridine carboxamide) in the presence or absence of visible light. A small increase in mutagenicity was observed with amsacrine but not with acridine carboxamide. A series of analogues of amsacrine were then tested, and a striking relationship was found between the minimum drug concentration for mutagenicity and DNA binding affinity. In each case, photoirradiation was associated with a small increase in mutagenicity. Each of the compounds showing the photo-enhancement effect was capable of reversible one-electron oxidation. It is suggested that this oxidation occurs in bacteria, and that the DNA binding constant of the resulting acridine radical species will increase because of the extra positive charge. This increased DNA binding would be sufficient to explain the photo-enhancement of mutagenicity of these drugs.

In vitro and in vivo mutagenicity studies on sporidesmin, the toxin associated with facial eczema in ruminants

Sporidesmin, a fungal toxin with widespread distribution within New Zealand, is thought to exert toxic effects through oxidative damage. The purified chemical was tested for its ability to cause point mutations in four strains of Salmonella typhimurium (TA98, TA100, TA102 and TA1537), in the presence and absence of exogenous metabolic activation. Although toxic effects were seen at concentrations exceeding 400 mu gl/plate, there were no significant increases in revertant colonies. In strain TA102, these results were not modified by the presence of glutathione. In AA8 Chinese hamster cells, sporidesmin acted as a potent clastogen, causing chromosomal breaks at concentrations as low as 3 ng/ml, where there was very little reduction in cell viability. Effects were primarily at the chromatid level, but some chromosomal events were also seen. Following low doses, the most common events were chromatid deletions and induction of double minute chromosomes. Interchange events occurred at concentrations of 10 ng/ml and above. The most common of these events was an incomplete chromatid interchange, although some examples of complete chromatid and chromosomal interchange were seen. These in vitro experiments were subsequently extended to an in vivo study of sporidesmin-induced lymphocytic micronuclei (MN) in sheep. In a double blind experiment, 5 sheep were treated with a single high dose of sporidesmin. Blood samples were taken from these, and from 5 untreated sheep, at various intervals before and after treatment. Peripheral blood lymphocytes cultures were harvested and scored for MN in cytokinesis-blocked cells, as a measure of clastogenic activity of sporidesmin in vivo. Following decoding, statistical analysis of the data revealed no significant differences between the MN levels in peripheral blood lymphocytes of sporidesmin-treated and untreated sheep. Although the possibility still exists that clastogenic effects could occur in other species, the data indicate that sporidesmin is not a clastogen in sheep, even though this species is highly susceptible to the toxic effects of sporidesmin.

Mutagenicity tests as a monitoring tool for potential mutagens and carcinogens in shellfish gathering areas of New Zealand

Abstract Shellfish may bioaccumulate a variety of chemicals, some of which are mutagenic or carcinogenic to humans. Mutagenicity tests provide an integrated way of detecting these chemicals. This paper describes the application of two such tests to New Zealand shellfish in laboratory and field situations. The bacterial mutagenicity test gave positive results on a nitric acid extract of green‐lipped mussels (Perna canaliculus) that had been exposed to model carcinogens under laboratory conditions. When applied to Pacific oysters (Crassostrea gigas) that had been sampled from four different sites in the Manukau Harbour, the same methods detected mutagenic activity which varied both by date and area sampled. The micronucleus assay gave a readily scored measure of chromosome damage in gill tissues in both mussels and oysters, but presented some practical problems in field studies. Our studies emphasise the need to sample within a short time interval, and the advantage of using a complementary package of bacte...

Mouse micronucleus assays of sporidesmin, the toxin associated with facial eczema in ruminants

Sporidesmin, a fungal toxin with widespread distribution within New Zealand, was previously shown to be a potent clastogen in Chinese hamster cells in vitro, but not in peripheral blood lymphocytes of sheep in vivo. In mice, massive oral doses led only to slight increases in micronucleus levels in the bone marrow, despite highly significant changes to other toxicological parameters. It would appear that the intact animal is protected in some fashion from the clastogenic effects of sporidesmin, although this substance must still be considered a potential human genotoxic agent.