Recent Advances in In Vivo Genotoxicity Testing: Prediction of Carcinogenic Potential Using Comet and Micronucleus Assay in Animal Models

Genotoxic events have been known as crucial step in the initiation of cancer. To assess the risk of cancer, genotoxicity assays, including comet, micronucleus (MN), chromosomal aberration, bacterial reverse, and sister chromatid exchange assay, can be performed. Compared with in vitro genotoxicity assay, in vivo genotoxicity assay has been used to verify in vitro assay result and definitely provide biological significance for certain organs or cell types. The comet assay can detect DNA strand breaks as markers of genotoxicity. Methods of the in vivo comet assay have been established by Japanese Center for the Validation of Alternative Methods (JaCVAM) validation studies depending on tissue and sample types. The MN can be initiated by segregation error and lagging acentric chromosome fragment. Methods of the in vivo MN assay have been established by Organization for Economic Co-operation and Development (OECD) test guidelines and many studies. Combining the in vivo comet and MN assay has been regarded as useful methodology for evaluating genetic damage, and it has been used in the assessment of potential carcinogenicity by complementarily presenting two distinct endpoints of the in vivo genotoxicity individual test. Few studies have investigated the quantitative relation between in vivo genotoxicity results and carcinogenicity. Extensive studies emphasizes that positive correlation is detectable. This review summarizes the results of the in vivo comet and MN assays that have investigated the genotoxicity of carcinogens as classified by the International Agency for Research on Cancer (IARC) carcinogenicity database. As a result, these genotoxicity data may provide meaningful information for the assessment of potential carcinogenicity and for implementation in the prevention of cancer.

[1]  Report of the JaCVAM initiative international pre-validation studies of the in vivo rodent alkaline Comet assay for the detection of genotoxic carcinogens , 2013 .

[2]  Olga Tcheremenskaia,et al.  The new ISSMIC database on in vivo micronucleus and its role in assessing genotoxicity testing strategies. , 2012, Mutagenesis.

[3]  H. van Steeg,et al.  Can carcinogenic potency be predicted from in vivo genotoxicity data? a meta‐analysis of historical data , 2011, Environmental and molecular mutagenesis.

[4]  G. Omurtag,et al.  Investigation of DNA damage in patients with colorectal cancer and their first degree relatives , 2011 .

[5]  M. Fenech,et al.  Micronuclei frequency in peripheral blood lymphocytes and cancer risk: evidence from human studies. , 2011, Mutagenesis.

[6]  M. Fenech,et al.  Molecular mechanisms of micronucleus, nucleoplasmic bridge and nuclear bud formation in mammalian and human cells. , 2011, Mutagenesis.

[7]  Andreas Czich,et al.  Collaborative study on fifteen compounds in the rat-liver Comet assay integrated into 2- and 4-week repeat-dose studies. , 2010, Mutation research.

[8]  G. Hovhannisyan.  Fluorescence in situ hybridization in combination with the comet assay and micronucleus test in genetic toxicology , 2010, Molecular Cytogenetics.

[9]  Q. Hasan,et al.  Assessment of genetic damage in inflammatory, precancerous, and cancerous pathologies of the esophagus using the comet assay. , 2010, Genetic testing and molecular biomarkers.

[10]  K. Witt,et al.  Dose-response assessment of four genotoxic chemicals in a combined mouse and rat micronucleus (MN) and Comet assay protocol. , 2010, The Journal of toxicological sciences.

[11]  Michael Fenech,et al.  THE LYMPHOCYTE CYTOKINESIS-BLOCK MICRONUCLEUS CYTOME ASSAY AND ITS APPLICATION IN RADIATION BIODOSIMETRY , 2010, Health physics.

[12]  Marie Z. Vasquez.  Combining the in vivo comet and micronucleus assays: a practical approach to genotoxicity testing and data interpretation , 2009, Mutagenesis.

[13]  Riccardo Crebelli,et al.  Assessment of in vivo genotoxicity of the rodent carcinogen furan: evaluation of DNA damage and induction of micronuclei in mouse splenocytes. , 2010, Mutagenesis.

[14]  M. Stratton,et al.  The cancer genome , 2009, Nature.

[15]  W. Ohyama,et al.  Detection of micronucleated cells and gene expression changes in glandular stomach of mice treated with stomach-targeted carcinogens. , 2008, Mutation research.

[16]  S. Bonassi,et al.  Micronuclei frequency in peripheral blood lymphocytes of cancer patients: a meta-analysis. , 2008, Mutation research.

[17]  R. Sandhir,et al.  Protective effects of vitamin E against atrazine-induced genotoxicity in rats. , 2008, Mutation research.

[18]  P. N. Rao,et al.  Cytogenetic biomarkers for human cancer. , 2008, Frontiers in bioscience : a journal and virtual library.

[19]  S. Bonassi,et al.  Validation of micronuclei frequency in peripheral blood lymphocytes as early cancer risk biomarker in a nested case-control study. , 2008, Mutation research.

[20]  David Kirkland,et al.  Evaluation of the ability of a battery of three in vitro genotoxicity tests to discriminate rodent carcinogens and non-carcinogens III. Appropriate follow-up testing in vivo. , 2005, Mutation research.

[21]  Albrecht Poth,et al.  Genetic toxicity assessment: employing the best science for human safety evaluation part IV: Recommendation of a working group of the Gesellschaft fuer Umwelt-Mutationsforschung (GUM) for a simple and straightforward approach to genotoxicity testing. , 2007, Toxicological sciences : an official journal of the Society of Toxicology.

[22]  M. Fenech.  Cytokinesis-block micronucleus cytome assay , 2007, Nature Protocols.

[23]  R. Jesudasan,et al.  Chromosome 11 aneusomy in esophageal cancers and precancerous lesions--an early event in neoplastic transformation: an interphase fluorescence in situ hybridization study from south India. , 2007, World journal of gastroenterology.

[24]  M. Fenech,et al.  An increased micronucleus frequency in peripheral blood lymphocytes predicts the risk of cancer in humans. , 2006, Carcinogenesis.

[25]  M. Kirsch‐Volders,et al.  Chromosomal changes: induction, detection methods and applicability in human biomonitoring. , 2006, Biochimie.

[26]  S. Pfuhler,et al.  The in vivo comet assay: use and status in genotoxicity testing. , 2005, Mutagenesis.

[27]  B. Mazmancı,et al.  Induction of micronuclei by lambda-cyhalothrin in Wistar rat bone marrow and gut epithelial cells. , 2005, Mutagenesis.

[28]  E. Dybing,et al.  Comparison of carcinogenic and in vivo genotoxic potency estimates. , 2005, Basic & clinical pharmacology & toxicology.

[29]  A. Jha.  Genotoxicological studies in aquatic organisms: an overview. , 2004, Mutation research.

[30]  Martin M. Schumacher,et al.  Use of the alkaline in vivo Comet assay for mechanistic genotoxicity investigations. , 2004, Mutagenesis.

[31]  Richard F. Lee,et al.  Use of the single cell gel electrophoresis/comet assay for detecting DNA damage in aquatic (marine and freshwater) animals. , 2003, Mutation research.

[32]  V. Garaj-vrhovac,et al.  The alkaline Comet assay as biomarker in assessment of DNA damage in medical personnel occupationally exposed to ionizing radiation. , 2003, Mutagenesis.

[33]  Trevelyan Square,et al.  Recommendations for conducting the in vivo alkaline Comet assay , 2003 .

[34]  A. Pruski,et al.  Marine invertebrate eco-genotoxicology: a methodological overview. , 2002, Mutagenesis.

[35]  Y. Sasaki,et al.  The comet assay with 8 mouse organs: results with 39 currently used food additives. , 2002, Mutation research.

[36]  A. Pruski,et al.  MARINE INVERTEBRATE ECOGENOTOXICOLOGY: A METHODOLOGICAL OVERVIEW , 2002 .

[37]  M. Fenech.  The in vitro micronucleus technique. , 2000, Mutation research.

[38]  M. Spitz,et al.  Gamma-radiation-induced single cell DNA damage as a measure of susceptibility to lung cancer: a preliminary report. , 2000, International journal of oncology.

[39]  F. A. Aly,et al.  In vivo and in vitro studies on the genotoxicity of cadmium chloride in mice , 2000, Journal of applied toxicology : JAT.

[40]  J. T. Macgregor,et al.  In vivo rodent erythrocyte micronucleus assay. II. Some aspects of protocol design including repeated treatments, integration with toxicity testing, and automated scoring , 2000, Environmental and molecular mutagenesis.

[41]  R. Tice,et al.  Single cell gel/comet assay: Guidelines for in vitro and in vivo genetic toxicology testing , 2000, Environmental and molecular mutagenesis.

[42]  M. Fenech,et al.  The HUman MicroNucleus Project--An international collaborative study on the use of the micronucleus technique for measuring DNA damage in humans. , 1999, Mutation research.

[43]  T. Orsière,et al.  Micronucleated lymphocyte rates from head-and-neck cancer patients. , 1999, Mutation research.

[44]  M. Hayashi,et al.  Evaluation of a tissue homogenization technique that isolates nuclei for the in vivo single cell gel electrophoresis (comet) assay: a collaborative study by five laboratories. , 1998, Mutation research.

[45]  C. C. Lombardi,et al.  Measurement and characterization of micronuclei in cultured primary lung cells of mice following inhalation exposure to benzene. , 1998, Mutagenesis.

[46]  E. Mereto,et al.  Increased frequency of micronucleated kidney cells in rats exposed to halogenated anaesthetics. , 1998, Mutation research.

[47]  Makoto Hayashi,et al.  Evaluation of the rat micronucleus test with bone marrow and peripheral blood: Summary of the 9th collaborative study by CSGMT/JEMS·MMS , 1998, Environmental and molecular mutagenesis.

[48]  R. Santella.  DNA Damage as an Intermediate Biomarker in Intervention Studies , 1997, Proceedings of the Society for Experimental Biology and Medicine. Society for Experimental Biology and Medicine.

[49]  Y. Sasaki,et al.  Simple detection of chemical mutagens by the alkaline single-cell gel electrophoresis (Comet) assay in multiple mouse organs (liver, lung, spleen, kidney, and bone marrow). , 1997, Mutation research.

[50]  Y. Sasaki,et al.  Detection of rodent liver carcinogen genotoxicity by the alkaline single-cell gel electrophoresis (Comet) assay in multiple mouse organs (liver, lung, spleen, kidney, and bone marrow). , 1997, Mutation research.

[51]  H. Shimada,et al.  An improved method for the mouse liver micronucleus test. , 1997, Mutation research.

[52]  T Sofuni,et al.  Evaluation of the rodent micronucleus assay in the screening of IARC carcinogens (groups 1, 2A and 2B) the summary report of the 6th collaborative study by CSGMT/JEMS MMS. Collaborative Study of the Micronucleus Group Test. Mammalian Mutagenicity Study Group. , 1997, Mutation research.

[53]  M. Dusinska,et al.  Comet assay in human biomonitoring studies: Reliability, validation, and applications , 1997, Environmental and molecular mutagenesis.

[54]  H E Poulsen,et al.  Benzene-induced genotoxicity in mice in vivo detected by the alkaline comet assay: reduction by CYP2E1 inhibition. , 1996, Mutation research.

[55]  E. Mirkova,et al.  Activities of the rodent carcinogens thioacetamide and acetamide in the mouse bone marrow micronucleus assay. , 1996, Mutation research.

[56]  P. Olive,et al.  The comet assay: a comprehensive review. , 1995, Mutation research.

[57]  D. Johnston,et al.  Sensitivity to genotoxic effects of bleomycin in humans: Possible relationship to environmental carcinogenesis , 1989, International journal of cancer.

[58]  G. Douglas,et al.  International Commission for Protection against Environmental Mutagens and Carcinogens. ICPEMC working paper No. 5. Genotoxicity tests as predictors of carcinogens: an analysis. , 1988, Mutation research.

[59]  R. Tice,et al.  A simple technique for quantitation of low levels of DNA damage in individual cells. , 1988, Experimental cell research.

[60]  J. Savage,et al.  A comment on the quantitative relationship between micronuclei and chromosomal aberrations. , 1988, Mutation research.

[61]  M. Shelby,et al.  Sister chromatid exchange analysis in lung and peripheral blood lymphocytes of mice exposed to methyl isocyanate by inhalation. , 1987, Environmental mutagenesis.

[62]  K J Johanson,et al.  Microelectrophoretic study of radiation-induced DNA damages in individual mammalian cells. , 1984, Biochemical and biophysical research communications.

[63]  J H Weisburger,et al.  Carcinogen testing: current problems and new approaches. , 1981, Science.

[64]  W Schmid,et al.  The micronucleus test. , 1975, Mutation research.