Mutation Research/Reviews in Genetic Toxicology最新文献

Results from short-term tests for carcinogens and our advanced knowledge on cellular and molecular mechanisms of carcinogenesis strongly suggest that carcinogens do not induce genetic changes necessarily by directly interacting with DNA. Therefore, it is not surprising to see that many carcinogens are not detectable by available genetic toxicology tests. Thus, it has become necessary to study nongenotoxic mechanisms of carcinogenesis and to provide methods to predict those carcinogens which escape from conventional mutation tests. One possible nongenotoxic mechanism of carcinogenesis which is supported by abundant experimental evidence is inhibition of gap junctional intercellular communication. Many, but not all, tumor-promoting agents have been shown to inhibit the communication of cultured cells as well as in vivo. Molecular mechanisms of gap junctional intercellular communication control revealed that connexin (gap junction) genes form a family of tumor suppressor genes. Control mechanisms of expression as well as function of connexins are vulnerable to various carcinogenic insults, notably to nongenetoxic carcinogens. Thus, studies on the role of connexins in cell growth and carcinogenesis may prove to be useful for establishing a mechanism-based test to detect certain types of nongenotoxic carcinogens.

The U.S. Environmental Protection Agency (USEPA) recently proposed new guidelines to update and replace the 1986 USEPA Guidelines for Carcinogen Risk Assessment. Today, there is a better understanding of the variety of modes by which carcinogens can operate that did not exist when the 1986 USEPA guidelines were published. Many laboratories are adding new test protocols in their programs directed at questions concerning the mechanisms of action of carcinogens. In response to the evolving science of carcinogenesis, the new guidelines provide an analytical framework for incorporating all relevant biological information and recognizing a variety of situations regarding cancer risk. In addition, the guidelines are flexible enough to allow consideration of future scientific advances.

Emerging short-term bioassays for chemically-induced carcinogenesis are dependent for their relevance to human risk assessment on the degree of coincidence of human and rodent tumor pathways. Since these pathways do not always converge, these new tests may have a number of unanticipated pitfalls. Models of liver and renal tumors are described. The results from Rb and p53 tumor suppressor gene transgenic animals are compared to human tumor syndromes. The question of mutagenic and epigenetic fingerprints of chemicals versus the cell-specific selection of spontaneous mutations is debated. Examples of specific pitfalls, such as the recently discovered Helicobacter hepaticus promoted liver tumor in mice are presented. The rat pseudogenes for p53 and the rare role of p53 in most important rodent tumor models other than epithelial tumors present experimental quandaries. The differential effects of carcinogens during various stages of rodent perinatal and adult development are also discussed. It is concluded that the pathways of both animal models and their human counterparts should be better identified so that realistic endpoint markers can be chosen for human carcinogenic risk assessment.

Benzodiazepines are a group of drugs which have been extensively used for their activities as an anti-anxiety, sedative, muscle relaxant and anti-convulsant. Benzodiazepines at present are the most commonly prescribed drugs. Some of these drugs are teratogenic and also carcinogenic in experimental animals. The wide human exposure to this group of drugs throughout the world is of great concern for human health. In the present review, we have attempted to evaluate and update the mutagenic and genotoxic effects of four of the most commonly used benzodiazepines, i.e., chlordiazepoxide (CDZ), diazepam (DZ), nitrazepam (NZ) and oxazepam (OZ) based on available literature.

Radon exposure has been linked to lung carcinogenesis in both human and animal studies. Studies of smoking and nonsmoking uranium miners indicate that radon alone is a risk factor for lung cancer at the levels encountered by these miners, although the possibility exists that other substances in the mine environment affect the radon-induced response. The relevance of data from mines to the lower-exposure home environment is often questioned; still, a recent study of miners exposed to relatively low radon concentrations demonstrated a statistically significant increase for lung and laryngeal cancer deaths. In two major series of experiments with rats, the primary carcinogenic effect found was respiratory tract tumors, and evidence for an inverse exposure-rate effect was also noted. Although this inverse dose-rate effect also has been described in underground miner studies, it may not similarly apply to radon in the home environment. This observation is due to the fact that, below a certain exposure, cells are hit once or not at all, and one would not expect any dose-rate effect, either normal or inverse. Because some chromosome aberrations persist in cycling cells as stable events, cytogenetic studies with radon are being performed to help complete the understanding of the events leading to radon-induced neoplasia. Radon has been found to induce 13 times as much cytogenetic damage (as measured by the occurrence of micronuclei) than a similar dose of ⁶⁰Co. A wide variety of mutation systems have demonstrated alpha-particle mutagenesis; recent investigations have focused on the molecular basis of alpha-induced mutagenesis. Gene mutations are induced by radon in a linear and dose-dependent fashion, and with a high biological effect relative to low-LET irradiation. Studies of the hprt locus show that approximately half of the alpha-induced mutations arise by complete deletion of the gene; the remaining mutations are split between partial deletions, rearrangements, and events not detectable by Southern blot or PCR exon analysis. Although other mutation systems do not show the same spectra as observed in the hprt gene (suggesting that the gene environment affects response), DNA deletions or multilocus lesions of various size appear to be predominant after radon exposure. As data emerge regarding radon-induced changes at the chromosomal and molecular level, the mechanisms involved in radon carcinogenesis are being clarified. This information should increase the understanding of risk at the low exposure levels typically found in the home.

The Collaborative Study Group for the Micronucleus Test (CSGMT) is one of the task groups in the Mammalian Mutagenesis Study Group (MMS) of the Environmental Mutagen Society of Japan (JEMS). It was established in 1982 and has made efforts to understand what the micronucleus test is, what are the advantages and disadvantages of the test as an in vivo detection system for mutagens/carcinogens, and to establish a standard protocol applicable to numerous chemicals. Members of the CSGMT have published more than 75 papers as part of collaborative studies and have contributed to the understanding of the nature of the micronucleus test and to setting guidelines for testing of medicinal and other chemicals. The CSGMT held some workshops to share up-to-date knowledge and techniques on the micronucleus test. Through workshops and collaborative studies, the CSGMT contributed to the maintaining of a high standard of knowledge and techniques among Japanese researchers of the micronucleus test. This paper reviews achievements made by the CSGMT until now.

Assessing urine mutagenicity with the Salmonella mutagenicity test is often limited by the volumes of the samples. Optimization of the assay was performed with factorial and Doehlert designs. Two fractional factorial designs 2^3-1 (3 factors, 4 experiments) were used to estimate the main effects of the percent S9 in the mix, the time of liquid incubation, the inoculum size and the growth conditions. A Doehlert design (3 factors, 13 experiments) was used to study the main effects and the interactions of the NADP, G6P and S9 in the mix. The positive markers were benzo[a]pyrene (BaP, 0.3 μg/plate) and a pool of smokers' urine (SU, 1.25 ml equivalent/plate). The response was limited to the induction factor (IF, number of induced revertants/number of spontaneous revertants) with Salmonella typhimurium TA98. The optimal conditions for BaP were: a 60 min period of liquid incubation and a volume of 0.1 ml (approx. 10⁸ cells/plate) of an overnight culture grown in 50 ml of Nutrient Broth No. 2 from a 250 ml flask. The S9 mix (0.1 ml, final volume) included 1.5% of S9, 1.0 mM NADP and 4.4 mM G6P. The maximal IF was 15.79. The optimal conditions for SU were: a 60 min period of liquid incubation and a volume of 0.1 ml (approx. 10⁸ cells/plate) of an overnight culture grown in 7 ml of Nutrient Broth No. 2 from a 20 × 180 mm tube. The S9 mix (0.1 ml, final volume) included: 4% S9, 4.2 mM NADP and 5.2 mM G6P. The maximal I7F was 10.95. These optimal conditions did not modify the spontaneous frequencies of the tester strains: TA97a, TA98, TA100 and TA102. The dose-response curves of mutagenic urine samples were found to be non-linear. This micromethod required 8-fold less urine sample and 12.5-fold less liver homogenate as compared to the standard plate incorporation assay and was from 6.2- to 11.8-fold more sensitive to evaluate urine mutagenicity. The sensitivity of this technique was found to be limited to individuals smoking more than approx. 5 cigarettes/day by the standard extraction-concentration procedure.

Mammalian cells are exposed to a wide variety of genotoxic stresses from both endogenous and exogenous sources. Cells typically exhibit cell cycle delays, or checkpoints, in response to acute genotoxic stress. Other types of cellular responses to DNA damage include apoptosis and probably increases in DNA repair levels. These response pathways are altered in cancer cells, by genetic alterations such as overexpression or mutation of oncogenes, or loss of tumor suppressor gene functions. As cancer chemotherapy relies primarily on the selective killing of cancer cells by DNA-damaging agents, genetic alterations affecting cellular stress response pathways may affect the outcome of cancer treatment.