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Environmental Science and Pollution Research

, Volume 24, Issue 23, pp 19267–19281 | Cite as

Effects of low doses of glyphosate on DNA damage, cell proliferation and oxidative stress in the HepG2 cell line

  • Vilena Kašuba
  • Mirta MilićEmail author
  • Ružica Rozgaj
  • Nevenka Kopjar
  • Marin Mladinić
  • Suzana Žunec
  • Ana Lucić Vrdoljak
  • Ivan Pavičić
  • Ana Marija Marjanović Čermak
  • Alica Pizent
  • Blanka Tariba Lovaković
  • Davor Želježić
Research Article

Abstract

We studied the toxic effects of glyphosate in vitro on HepG2 cells exposed for 4 and 24 h to low glyphosate concentrations likely to be encountered in occupational and residential exposures [the acceptable daily intake (ADI; 0.5 μg/mL), residential exposure level (REL; 2.91 μg/mL) and occupational exposure level (OEL; 3.5 μg/mL)]. The assessments were performed using biomarkers of oxidative stress, CCK-8 colorimetric assay for cell proliferation, alkaline comet assay and cytokinesis-block micronucleus (CBMN) cytome assay. The results obtained indicated effects on cell proliferation, both at 4 and 24 h. The levels of primary DNA damage after 4-h exposure were lower in treated vs. control samples, but were not significantly changed after 24 h. Using the CBMN assay, we found a significantly higher number of MN and nuclear buds at ADI and REL after 4 h and a lower number of MN after 24 h. The obtained results revealed significant oxidative damage. Four-hour exposure resulted in significant decrease at ADI [lipid peroxidation and glutathione peroxidase (GSH-Px)] and OEL [lipid peroxidation and level of total antioxidant capacity (TAC)], and 24-h exposure in significant decrease at OEL (TAC and GSH-Px). No significant effects were observed for the level of reactive oxygen species (ROS) and glutathione (GSH) for both treatment, and for 24 h for lipid peroxidation. Taken together, the elevated levels of cytogenetic damage found by the CBMN assay and the mechanisms of primary DNA damage should be further clarified, considering that the comet assay results indicate possible cross-linking or DNA adduct formation.

Keywords

Glyphosate HepG2 Genotoxicity Comet assay Cytotoxicity Low doses Oxidative damage Cytochalasin B-blocked micronucleus assay 

Notes

Acknowledgments

This work was financially supported by Project No. 8366 Organic Pollutants in Environment—Markers and Biomarkers of Toxicity (OPENTOX), funded by the Croatian Science Foundation.

Compliance with ethical standard

Conflict of interest

The authors declare that they have no conflict of interest.

Supplementary material

11356_2017_9438_MOESM1_ESM.docx (15 kb)
Supplementary Table 1 (DOCX 15 kb).

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Copyright information

© Springer-Verlag GmbH Germany 2017

Authors and Affiliations

  • Vilena Kašuba
    • 1
  • Mirta Milić
    • 1
    Email author
  • Ružica Rozgaj
    • 1
  • Nevenka Kopjar
    • 1
  • Marin Mladinić
    • 1
    • 2
  • Suzana Žunec
    • 3
  • Ana Lucić Vrdoljak
    • 3
  • Ivan Pavičić
    • 4
  • Ana Marija Marjanović Čermak
    • 4
  • Alica Pizent
    • 5
  • Blanka Tariba Lovaković
    • 5
  • Davor Želježić
    • 1
  1. 1.Mutagenesis UnitInstitute for Medical Research and Occupational HealthZagrebCroatia
  2. 2.Xellia Ltd.ZagrebCroatia
  3. 3.Toxicology UnitInstitute for Medical Research and Occupational HealthZagrebCroatia
  4. 4.Radiation Dosimetry and Radiobiology UnitInstitute for Medical Research and Occupational HealthZagrebCroatia
  5. 5.Analytical Toxicology and Mineral Metabolism UnitInstitute for Medical Research and Occupational HealthZagrebCroatia

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