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Formation of micronuclei, nuclear buds and nucleoplasmic bridges in porcine kidney PK15 cells after individual and combined exposure to fumonisin B1, beauvericin and ochratoxin A

The genotoxic potential of FB1, BEA and OTA as well as their combinations, has been investigated by means of an in vitro micronucleus assay on porcine kidney PK15 cells. Cells were treated with 0.05, 0.5 and 5 ug/ml of mycotoxin alone or two- or three-mycotoxin combination for 24 and 48 h. Mytomycin C (0.03 uM) and colchicine (1.20 uM) were used as positive controls. 1000 binucleated cells (7% Giemsa) were scored for the presence of micronucleus (MN), nuclear buds (NB) and nucleoplasmic bridges (NPB). OTA (0.05 ug/ml) significantly increased frequency of MN (13‰ ) in respect to control (4‰ ). 0.5 and 5 ug/ml of each mycotoxin equally increased the MN frequency (about 15‰ and 19‰ , respectively). The two and three-mycotoxin treatment increased the frequency of MN, mostly in additive manner. OTA (0.5 ug/ml) and BEA (5 ug/ml) significantly increased the frequency of NPB (8-10‰ ), while FB1 did not significantly affect the formation of NPB in any of concentrations. Frequency of nuclear buds was significantly increased after treatment with 5 ug/ml of each mycotoxin (88 to 95‰ ), as compared to control (64‰ ). However, combined treatment resulted in decrease of nuclear buds, especially after 48 h of exposure, which was probably connected with extrusion of micronuclei. All of three mycotoxins were predominantly clastogenic. In conclusion, this study provided additional evidence for the hypothesis that OTA and FB1 are carcinogens with genotoxic properties. To our knowledge, the present study is the first report on induction of clastogenic effects of BEA in cell cultures.

fumonisin B1; beauvericin; ochratoxin A; genotoxicity; carcinogenicity; micronucleus; nuclear buds; nucleoplasmic bridges; clastogen; mytomicin; colchicines; kidney cells.

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