Cytotoxicity and genotoxicity of anticancer drug residues and their mixtures in experimental model with zebrafish liver cells

- Cyclophosphamide and ifosfamide induce genotoxic effects at high concentrations in ZFL cells.
- Mixture of anticancer drugs induces DNA damage at environmentally relevant concentrations.
- DNA damaging potential of the mixture was higher than that of single drugs.
- Mixtures of anticancer drugs may pose threat to aquatic organisms.

Anticancer drugs enter aquatic environment predominantly via hospital and municipal wastewater effluents where they may, due to their genotoxic potential, cause adverse environmental effects even at very low doses. In this study we evaluated cytotoxic and genotoxic potential of two widely used anticancer drugs, cyclophosphamide (CP) and ifosfamide (IF) as individual compounds and in a complex mixture together with 5-fluorouracil (5-FU) and cisplatin (CDDP) because these four drugs have been frequently detected in an oncological ward effluents. As an experimental model we used zebrafish liver cell (ZFL) line. The cytotoxicity was determined with the MTS assay and genotoxicity with the comet assay and cytokinesis block micronucleus (CBMN) assay that measure the formation of DNA strand breaks and genomic instability, respectively. CP and IF exerted low cytotoxicity towards ZFL cells. Both compounds induced DNA strand breaks and genomic instability, however at relatively high concentrations that are not relevant for the contamination of aquatic environment. The mixture of CP, IF, 5-FU and CDDP was tested at maximal detected concentrations of each drug as determined in the effluents from the oncological ward. The mixture was not cytotoxic and did not induce genomic instability, but it induced significant increase in the formation of DNA strand breaks at concentrations of individual compounds that were several orders of magnitude lower from those that were effective when tested as individual compounds. The results indicate that such mixtures of anticancer drugs may pose a threat to aquatic organisms at environmentally relevant concentrations and contribute to the accumulating evidence that it is not always possible to predict adverse effects of complex mixtures based on the toxicological data for individual compounds.

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