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Genotoxic potential of selected cytostatic drugs in human and zebrafish cells

  • Fate and effects of the residues of anticancer drugs in the environment
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Abstract

Due to their increasing use, the residues of anti-neoplastic drugs have become emerging pollutants in aquatic environments. Most of them directly or indirectly interfere with the cell’s genome, which classifies them into a group of particularly dangerous compounds. The aim of the present study was to conduct a comparative in vitro toxicological characterisation of three commonly used cytostatics with different mechanisms of action (5-fluorouracil [5-FU], cisplatin [CDDP] and etoposide [ET]) towards zebrafish liver (ZFL) cell line, human hepatoma (HepG2) cells and human peripheral blood lymphocytes (HPBLs). Cytotoxicity was determined by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay and acridine orange/ethidium bromide staining. All three drugs induced time- and dose-dependent decreases in cell viability. The sensitivity of ZFL and HepG2 cells towards the cytotoxicity of 5-FU was comparable (half maximal inhibitory concentration (IC50) 5.3 to 10.4 μg/mL). ZFL cells were more sensitive towards ET- (IC50 0.4 μg/mL) and HepG2 towards CDDP- (IC50 1.4 μg/mL) induced cytotoxicity. Genotoxicity was determined by comet assay and cytokinesis block micronucleus (CBMN) assay. ZFL cells were the most sensitive, and HPBLs were the least sensitive. In ZFL cells, induction of DNA strand breaks was a more sensitive genotoxicity endpoint than micronuclei (MNi) induction; the lowest effective concentration (LOEC) for DNA strand break induction was 0.001 μg/mL for ET, 0.01 μg/mL for 5-FU and 0.1 μg/mL for CDDP. In HepG2 cells, MNi induction was a more sensitive genotoxicity endpoint. The LOEC values were 0.01 μg/mL for ET, 0.1 μg/mL for 5-FU and 1 μg/mL for CDDP. The higher sensitivity of ZFL cells to cytostatic drugs raises the question of the impact of such compounds in aquatic ecosystem. Since little is known on the effect of such drugs on aquatic organisms, our results demonstrate that ZFL cells provide a relevant and sensitive tool to screen genotoxic potential of environmental pollutant in the frame of hazard assessment.

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Acknowledgments

This study received funding from the Seventh Framework Programme FP7/2007-2013 under grant agreement No. 265264, from the Ministry of Science, Education and Sports of the Republic of Croatia under grant agreement No. 022-0222148-2125, from the Slovenian Research Agency: Program P1-0245 and Program of bilateral collaboration between Croatia and Slovenia (Grant Nos. 533-19-14-0003 (Croatia) and BI-HR/14-15-004 (Slovenia)).

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Authors and Affiliations

  1. Mutagenesis Unit, Institute for Medical Research and Occupational Health, Ksaverska cesta 2, 10000, Zagreb, Croatia

    Goran Gajski, Marko Gerić & Vera Garaj-Vrhovac

  2. Department of Genetic Toxicology and Cancer Biology, National Institute of Biology, Večna pot 111, 1000, Ljubljana, Slovenia

    Bojana Žegura, Matjaž Novak, Jana Nunić, Džejla Bajrektarević & Metka Filipič

  3. Ecological Engineering Institute, Ljubljanska ulica 9, 2000, Maribor, Slovenia

    Matjaž Novak

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  1. Goran Gajski
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  2. Marko Gerić
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Correspondence to Metka Filipič.

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Gajski, G., Gerić, M., Žegura, B. et al. Genotoxic potential of selected cytostatic drugs in human and zebrafish cells. Environ Sci Pollut Res 23, 14739–14750 (2016). https://doi.org/10.1007/s11356-015-4592-6

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