Giromini Carlotta, Rebucci Raffaella, Fusi Eleonora, Rossi Luciana, Saccone Francesca, Baldi Antonella
Department of Health, Animal Science and Food Safety, Università degli Studi di Milano, Via D. Trentacoste 2, 20134, Milan, Italy.
Cell Biol Toxicol. 2016 Jun;32(3):249-58. doi: 10.1007/s10565-016-9332-2. Epub 2016 May 6.
This study aimed to investigate the in vitro damage induced by ochratoxin A (OTA) in BME-UV1 and MDCK epithelial cells. Both cells lines were treated with OTA (0 up to 10 μg/mL), and cell viability (MTT assay), membrane stability (lactate dehydrogenase (LDH) release assay) and apoptotic cell rate (Tunel assay) were investigated. Further, the effect of the incubation with OTA has been evaluated at DNA level by the determination of DNA integrity, by the quantification of DNA adduct formation (8-hydroxy-2'-deoxyguanosine (8-OHdG)) and by the assessment of the global DNA methylation status (5-methyl-cytosine (5-mC)). The obtained results showed that after 24 h of OTA treatment, BME-UV1 cell viability was reduced in a dose-dependent way. OTA significantly (P < 0.05) increased LDH release in BME-UV1 cells at all concentrations tested. OTA (1.25 μg/mL) induced 35 % LDH release in MDCK cells (P < 0.05). A significant (P < 0.05) change in percentages of apoptotic BME-UV1 (10 ± 0.86) and MDCK (25 ± 0.88) cells was calculated when the cells were co-incubated with OTA. The level of 8-OHdG adduct formation was significantly (P < 0.05) increased in BME-UV1 cells treated with 1.25 μg/mL of OTA. The results of the present study suggest that a different mechanism of action may occur in these cell lines. Graphical abstract Study results overview.
本研究旨在调查赭曲霉毒素A（OTA）对BME-UV1和MDCK上皮细胞的体外损伤。两种细胞系均用OTA（0至10μg/mL）处理，并研究细胞活力（MTT法）、膜稳定性（乳酸脱氢酶（LDH）释放法）和凋亡细胞率（Tunel法）。此外，通过测定DNA完整性、定量DNA加合物形成（8-羟基-2'-脱氧鸟苷（8-OHdG））以及评估整体DNA甲基化状态（5-甲基胞嘧啶（5-mC）），在DNA水平上评估了与OTA共孵育的效果。所得结果表明，OTA处理24小时后，BME-UV1细胞活力呈剂量依赖性降低。在所有测试浓度下，OTA均显著（P<0.05）增加了BME-UV1细胞中的LDH释放。OTA（1.25μg/mL）在MDCK细胞中诱导了35%的LDH释放（P<0.05）。当细胞与OTA共孵育时，计算得出凋亡的BME-UV1细胞（10±0.86）和MDCK细胞（25±0.88）的百分比有显著（P<0.05）变化。在用1.25μg/mL OTA处理的BME-UV1细胞中，8-OHdG加合物形成水平显著（P<0.05）增加。本研究结果表明，这些细胞系可能存在不同的作用机制。图形摘要 研究结果概述。
