Abstract
Atrazine, one of the herbicide, is classified as a possible human carcinogen that may cause breast and ovarian cancers. The mutagenic and genotoxic properties of atrazine have been studied using a variety of assays in the past but the results were contradictory. This fact indicates that a more comprehensive assessment of the potential genotoxicity of atrazine is needed. For this reasons, we investigated the mutant frequency on lacI gene and induction of binucleated micronuclei formation by atrazine in vitro using the Big Blue® Rat2 lacI transgenic fibroblasts, which are transfected by a lambda shuttle vector. These assays carried out at the concentrations of 261.7, 130.8 and 65.4 μg/mL of atrazine, the highest concentration giving 80% survival. These two endpoints were used to compare the induction of point mutations with cytogenetic damages by atrazine under same exposure condition. No significant changes of mutant frequency and binucleated micronuclei frequency induced by atrazine were observed both in the transgenic mutagenesis assay and cytokinesis blocking assay in the absence of metabolic activation system using the Big Blue® Rat2 lacI transgenic fibroblasts. These results suggest that atrazine did not induce mutation and cytogenetic damage in this experimental system.
This is a preview of subscription content, log in via an institution to check access.
Similar content being viewed by others
References
Selim, H. Retention and runoff losses of atrazine and metribuzin in soil. J. Environ. Qual. 32, 1058–1071 (2003).
Pogrmic-Majkic, K., Fa, S., Dakic, V., Kaisarevic, S. & Kovacevic, R. Upregulation of peripubertal rat Leydig cell steroidogenesis following 24 h in vitro and in vivo exposure to atrazine. Toxicol. Sci. 118, 52–60 (2010).
Hovey, R. C. et al. Quantitative assessment of mammary gland development in female Long Evans rats following in utero exposure to atrazine. Toxicol. Sci. 119, 380–390 (2011).
Clements, C., Ralph, S. & Petras, M. Genotoxicity of select herbicides in Rana catesbeiana tadpoles using the alkaline single-cell gel DNA electrophoresis (comet) assay. Environ. Mol. Mutagen 29, 277–288 (1997).
Kligerman, A. D., Doerr, C. L., Tennant, A. H. & Peng, B. Cytogenetic studies of three atrazine herbicides 11 In vivo micronucleus studies in mouse bone marrow. Mutat. Res. 471, 107–112 (2000).
Garaj-Vrhovac, V. & Zeljezic, D. Evaluation of DNA damage in workers occupationally exposed to pesticides using single-cell gel electrophoresis (SCGE) assay, pesticide genotoxicity revealed by comet assay. Mutat. Res. 469, 279–285 (2000).
Nwani, C. D. et al. Mutagenic and genotoxic assessment of atrazine-based herbicide to freshwater fish Channa punctatus (Bloch) using micronucleus test and single cell gel electrophoresis. Environ. Toxicol. Pharmacol. 31, 314–322 (2011).
Dycaico, M. J. et al. The use of shuttle vectors for mutation analysis in transgenic mice and rats. Mutat. Res. 307, 461–478 (1994).
Gorelick, N. J. Overview of mutation assays in transgenic mice for routine testing. Environ. Mol. Mutagen 25, 218–230 (1995).
Lundberg, K. S., Provost, G. S., Kretz, P. L. & Short, J. M. The use of selection in recovery of transgenic targets for mutation analysis. Mutat. Res. 301, 99–105 (1993).
Heddle, J. A. & Tao, K. S. The transmission rate of the lacI transgene from the Big Blue™ mouse. Mutat. Res. 348, 63–66 (1995).
Kohler, S. W. et al. Spectra of spontaneous and mutagen-induced mutations in the lacI gene in transgenic mice. Proc. Natl. Acad. Sci. USA 88, 7958–7962 (1991).
Tao, K. S., Urlando, C. & Heddle, J. A. Comparison of somatic mutation in a transgenic versus host locus. Proc. Natl. Acad. Sci. USA 90, 10681–10685 (1993).
Ryu, J. C. et al. Mutation spectrum of 4-nitroquinoline-N-oxide in the lacI transgenic Big Blue® Rat2 cell line. Mutat. Res. 445, 127–135 (1999).
Ryu, J. C., Kim, Y. J. & Chai, Y. G. Mutation spectrum of 1,2-dibromo-3-chloropropane, an endocrine disruptor, in the lacI transgenic Big Blue Rat2 fibroblast cell line. Mutagenesis 17, 301–307 (2002).
Besplug, J., Filkowski, J., Burke, P., Kovalchuk, I. & Kovalchuk, O. Atrazine induces homologous recombination but not point mutation in the transgenic plantbased biomonitoring assay. Arch. Environ. Contam. Toxicol. 46, 296–300 (2004).
Cavas, T. In vivo genotoxicity evaluation of atrazine and atrazine-based herbicide on fish Carassius auratus using the micronucleus test and the comet assay. Food Chem. Toxicol. 49, 1431–1435 (2011).
Campos-Pereira, F. D. et al. Early cytotoxic and genotoxic effects of atrazine on Wistar rat liver: a morphological, immunohistochemical, biochemical, and molecular study. Ecotoxicol. Environ. Saf. 78, 170–177 (2012).
Ribas, G. et al. Lack of genotoxicity of the herbicide atrazine in cultured human lymphocytes. Mutat. Res. 416, 93–99 (1998).
al-Sabti, K. & Metcalfe, C. D. Fish micronuclei for assessing genotoxicity in water. Mutat. Res. 343, 121–135 (1995).
Inboot, W. et al. Genotoxicity and antigenotoxicity of the methanol extract of Cleistocalyx nervosum var. paniala seed using a Salmonella mutation assay and rat liver micronucleus tests. Mol. Cell. Tox. 8, 19–24 (2012).
An, Y. R. et al. Genomic comparison of insecticides and herbicide in human hepatoma (HepG2) cell line. Mol. Cell. Toxicol. 6, 381–386 (2010).
Zeljezic, D., Garaj-Vrhovac, V. & Perkovic, P. Evaluation of DNA damage induced by atrazine and atrazinebased herbicide in human lymphocytes in vitro using a comet and DNA diffusion assay. Toxicol. In Vitro 20, 923–935 (2006).
Author information
Authors and Affiliations
Corresponding author
Rights and permissions
About this article
Cite this article
Kim, YJ., Ryu, JC. Genotoxicity assessment of atrazine in the Big Blue® Rat2 lacI transgenic cell line. Toxicol. Environ. Health Sci. 4, 161–166 (2012). https://doi.org/10.1007/s13530-012-0131-8
Received:
Revised:
Accepted:
Published:
Issue Date:
DOI: https://doi.org/10.1007/s13530-012-0131-8