Impact of in vivo and in vitro exposure to selected cytostatics on DNA damage in haemocytes of freshwater mussels Unio pictorum and Unio tumidus

Abstract

The impact of 5-Fluorouracil (5-FU), Cisplatin (CP) and Etoposide (Eto) on DNA damage level was studied in vivo and in vitro on haemocytes of freshwater mussels Unio pictorum and U. tumidus using alkaline comet assay. For in vitro treatment, two different approaches were applied: i) isolated haemocytes were treated for 30 min in physiological solution, ii) primary culture of haemocytes was treated for 22h. For in vivo experiment, mussels were exposed in static system for 72h. CdCl2 was used as positive control. The level of DNA damage was analyzed by Comet Assay IV software. CdCl2 induced significant increase in DNA damage in vitro and in vivo. Increase of damage was also detected with Eto in vivo and in primary cultures of haemocytes (in vitro). For 5–FU, the significant increase of DNA damage was detected only in vivo, while CP did not induced increase of DNA damage in vivo or in vitro. Lack of genotoxic effects of cytostatics in isolated haemocytes can be attributed to short period of exposure (30 min), while the lack of the effects of 5-FU and CP in primary cultures can be assigned to the mechanisms of action of these drugs.