DNA damage induced by mutagens in plant and human cell nuclei in acellular comet assay.
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AbstractHigher plant cells have a long tradition of use in the studies on environmental mutagenesis in situ, especially in relation to human health risk determination. The studies on the response of plant and human cells to physical and chemical mutagens showed differences in their sensitivity. The differences in the presence of cell components in plants and humans could influence such response. Additionally, the level of the organization of the employed material could influence DNA-damaging effect: leukocytes are isolated cells and plant--an intact organism. To preclude these obstacles, the effects of direct treatment of isolated nuclei with genotoxic agents were determined to compare the sensitivity of plant and human cells. In the present study, we have determined the DNA-damaging effects of two chemical mutagens: maleic acid hydrazide (MH) and N-methyl-N-nitroso-urea (MNU) applied to isolated nuclei of both plant and human cells. In order to compare the sensitivity of the nuclei of Nicotiana tabacum var. xanthi and the nuclei of leukocytes, the acellular Comet assay was carried out. The results showed higher sensitivity of the nuclei of leukocytes as compared to the nuclei of plant cells to mutagenic treatment with the applied doses of MH and MNU.
CitationFolia Histochem. Cytobiol. 2006, 44 (2):127-131
JournalFolia Histochemica et Cytobiologica / Polish Academy of Sciences, Polish Histochemical and Cytochemical Society
DescriptionBiomarkers of exposure & effect: method development & validation Biomarker: DNA damageExposure/effect represented: maleic acid hydrazide (MH); N-methyl-N-nitroso-urea (MNU) / DNA damageTissue/biological material/sample size: isolated nuclei of double heterozygous Nicotiana tabacum var. xanthi; isolated nuclei of human leukocytes obtained from a healthy donorMethod of analysis: acellular comet assayQuality control: Each experiment was repeated twice and then three slides were analyzed per each experimental group. From the repeated experiments, the average values ▒SD of each parameter for each treatment group were calculated from the median values from each slide. The differences between the two groups were statistically evaluated by Student's t- test.Impact on outcome (including dose-response):the results demonstrate higher sensitivity of leukocyte nuclei than plant cell nuclei, especially using higher doses of the applied mutagens. Both mutagens applied to isolated nuclei induced DNA damage, and the strongest effect was observed in the case of 1 mM MNU. The treatment of isolated nuclei by the applied mutagens caused a dose-dependent effect, showed by different parameters of Dtudent's t- test. KEYWORDS - CLASIFFICATION: analysis;biomarkers of exposure & effect: method development & validation;cytology;Cell Nucleus;Comet Assay;drug effects;DNA Damage;genetics;Humans;Leukocytes;methods;Mutagens;pharmacology;Poland;Tobacco;validation;
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