Effects of trabectedin in the zebrafish Danio rerio: from cells to larvae

Abstract

Anticancer agents pose a great environmental risk due to their high toxicity. The aim of the current study is to assess the toxicity of trabectedin, a cytotoxic but atypical DNA binder, to liver cell line (ZFL) and embryo-larvae of the zebrafish Danio rerio employing an innovative approach. In ZFL cells, trabectedin cytotoxicity was measured using MTT and Trypan blue exclusion assay, and cell morphology was evaluated by fluorescence-activated cell sorting (FACS) and by immunofluorescence analysis. Trabectedin was 60-fold more toxic to ZFL cells than to zebrafish embryo-larvae in terms of mortality/cell viability, with mortality being observed in 42.7 µg.L−1 for embryo-larvae and non-viability in 0.04 µg.L−1 for cultured cells. Immunofluorescence staining showed morphology alterations of ZFL-cells exposed to trabectedin in a dose-dependent manner, from 0.04 to 0.15 µg.L−1. Furthermore, trabectedin induced morphological abnormalities to zebrafish embryo-larvae, such as tail malformations, pericardial edema and lack of equilibrium at concentrations lower than 50.3 µg.L−1. Regarding larvae behavior analysis, trabectedin increased velocity and total distance covered by zebrafish exposed to 42.7 µg.L−1 under dark conditions. These results reveal trabectedin to be toxic in both in vitro and in vivo zebrafish models, and thus the occurrence and persistence of this anticancer agent in the environment may represent a potential risk factor to the biota.

Keywords

• Anticancer agent
• Zebrafish locomotor behavior
• ZFL cells
• Developmental toxicity
• Cytotoxicity