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Assessment of DNA damage,cytotoxicity, and apoptosis in human hepatoma (HepG2) cells after flurochloridone herbicide exposure
Affiliation:1. Laboratorio de Ecotoxicología y Contaminación Ambiental, Instituto de, Investigaciones Marinas y Costeras (IIMyC), Facultad de Ciencias Exactas y, Naturales, Universidad Nacional de Mar del Plata (UNMdP), Consejo Nacional de Investigaciones Científicas y Técnicas (CONICET), D. Funes 3350, Mar del Plata 7600, Argentina;2. Instituto Nacional de Tecnología Agropecuaria INTA EEA Balcarce, Ruta Nacional 226, Km 73,5, Balcarce CP 7620, Buenos Aires, Argentina;3. CIMA-Facultad de Ciencias Exactas-UNLP, Argentina;4. Instituto de Ciencia y Tecnología de Alimentos Córdoba, Argentina;1. Department of Environmental and Molecular Toxicology, Oregon State University, Corvallis, OR, United States;2. School of Chemical, Biological and Environmental Engineering, Oregon State University, Corvallis, OR, United States;3. Oregon Nanoscience and Microtechnologies Institute, Eugene, OR, United States
Abstract:In vitro effects of flurochloridone (FLC) and its formulations Twin Pack Gold® [25% active ingredient (a.i.)] and Rainbow® (25% a.i.) were evaluated in HepG2 cells. Whereas cytokinesis-blocked micronucleus cytome (CBMN-cyt) and single-cell gel electrophoresis (SCGE) assays were employed for genotoxicity, MTT, neutral red, and apoptosis detections were used for cytotoxicity evaluation. Activities were tested within the concentration range of 0.25–15 μg/ml FLC. Results demonstrated that neither FLC nor Rainbow® was able to induce MNs. On the other hand, 5 μg/ml Twin Pack Gold® only increased MN frequency. Furthermore, 10 and 15 μg/ml of both formulations resulted in cellular cytotoxicity demonstrated by alterations in the nuclear division index and cellular death. A marked increase in the genetic damage index was observed after treatment with all compounds. SCGE assay appeared to be more sensitive bioassay for detecting primary DNA strand breaks at lower concentrations of FLC than did MN. Our results reveal that FLC and its two formulations trigger apoptosis on HepG2 cells. The results represent the first experimental evidence of the in vitro apoptogenic role exerted on mammalian cells by FLC and the FLC-based formulations Rainbow® and Twin Pack Gold®, at least on HepG2 cells.
Keywords:DNA single-strand breaks  HepG2 cells  Commercial herbicide formulations  Cytokinesis-blocked micronucleus cytome (CBMN-cyt) assay
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