Naslov
A potential role of calcium in apoptosis and aberrant chromatin forms in porcine kidney PK15 cells induced by individual and combined ochratoxin A and citrinin

Autori
Šegvić Klarić, Maja ; Želježić, Davor ; Rumora, Lada ; Peraica, Maja ; Pepeljnjak, Stjepan ; Domijan, Ana-Marija

Izvornik
Archives of toxicology (0340-5761) 86 (2012), 1; 97-107

Vrsta, podvrsta i kategorija rada
Radovi u časopisima, članak, znanstveni

Ključne riječi
nephrotoxins; genotoxicity; necrosis; BAPTA-AM; mycotoxin synergism

Sažetak
The aim of this study was to establish the involvement of calcium signalling in genotoxicity, apoptosis and necrosis evoked by ochratoxin A (OTA) and citrinin (CTN) alone or in combination in porcine kidney PK15 cells. Cell proliferation test (MTT) and trypan blue assays (24 h) demonstrated that CTN (IC50=73.5±1.0, 75.4±1.4 M, respectively) was less toxic than OTA (IC50=14.0±2.4, 20.5±1.0 M, respectively). To test their cytotoxic interactions two doses of single OTA (6 and 10 M) and CTN (30 and 50 M) and their combinations were applied. Combined treatment showed additive cytotoxic effects. OTA and CTN induced dose-depended increase in cytosolic calcium level (assessed with Fura-2 AM). However, combined treatment did not provoke additional increase in calcium signal. The rate of apoptosis and necrosis (DAPI-antifade staining) was significantly higher after 12 h than 24 h, while the frequencies of micronuclei (MNs) and nuclear buds (NBs) were higher after 24 h than 12 h treatment. Combined exposure resulted in apoptotic and necrotic synergism, while genotoxic effects of OTA+CTN were noted as antagonistic or additive. Co-exposure of cells to calcium chelator BAPTA-AM significantly reduced CTN and OTA+CTN- evoked apoptosis. 24 h after co-exposure to BAPTA- AM and a single OTA and CTN, MNs significantly decreased while NBs dropped significantly after co-treatment with BAPTA-AM and OTA+CTN. In conclusion, disturbance of Ca2+ homeostasis caused by OTA and CTN plays a significant role in cell genotoxicity and death.

Izvorni jezik
Engleski

Znanstvena područja
Biologija, Farmacija

POVEZANOST RADA