Comparative study of deoxynivalenol and 15-acetyldeoxynivalenol on gene expression in the human cell line Caco-2

To screen cellular effects on Caco-2 cells by deoxynivalenol (DON) and 15-acetyldeoxynivalenol (15ADON), toxin-treated Caco-2 cells were analyzed by DNA microarray. Exposure to either toxin induced up- or downregulation of genes in Caco-2 cells. Upregulation of 1290 and 3238 genes was observed for the DON- and 15ADON-treated groups, respectively, after a 60-min incubation period. This represented a greater than 1.5-fold change relative to the 0-min exposure (control) group. Five hundred and sixty-one genes showed a 1.5-fold upregulation in the both the DON- and 15ADON-treated groups. Similarly, 13 genes and 105 genes were upregulated with a log2 ratio > 2 (4-fold change) in the DON- and 15ADON-treated groups, respectively. The most remarkable gene upregulation corresponded to the inflammatory chemokine, IL-8, which showed a greater than 4-fold upregulation in response to both treatments. Deoxynivalenol and 15-acetyldeoxynivalenol at concentrations of 1 ug/mL were added separately to the AP chamber, and cells were harvested with 0.5% trypsin-EDTA after incubation for 0, 5, 30, and 60 min. Toxin-treated Caco-2 cells were analyzed by DNA microarray to screen for changes in gene expression in response to deoxynivalenol or 15-acetyldeoxynivalenol.

为探究脱氧雪腐镰刀菌烯醇（deoxynivalenol, DON）与15-乙酰脱氧雪腐镰刀菌烯醇（15-acetyldeoxynivalenol, 15ADON）对Caco-2细胞的细胞效应，研究人员采用DNA微阵列（DNA microarray）技术对毒素处理后的Caco-2细胞进行了分析。两种毒素单独处理均可诱导Caco-2细胞内基因的上调或下调表达。孵育60分钟后，DON处理组与15ADON处理组分别观测到1290个与3238个基因发生上调，该表达变化相较于0分钟（未处理对照）组均超过1.5倍。另有561个基因在DON与15ADON两个处理组中均出现1.5倍以上的上调表达。类似地，DON处理组与15ADON处理组中，分别有13个与105个基因的log₂比值大于2（即表达变化幅度达4倍以上）。其中上调幅度最显著的基因为炎症趋化因子IL-8，经两种毒素处理后其表达上调幅度均超过4倍。本实验采用1 μg/mL浓度的脱氧雪腐镰刀菌烯醇与15-乙酰脱氧雪腐镰刀菌烯醇，分别添加至AP侧室；分别于孵育0、5、30、60分钟后，使用0.5%胰蛋白酶-EDTA消化收集细胞。后续通过DNA微阵列技术分析毒素处理后的Caco-2细胞，以筛选脱氧雪腐镰刀菌烯醇或15-乙酰脱氧雪腐镰刀菌烯醇诱导的基因表达变化。