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In vitro effect of deoxynivalenol on the differentiation of human colonic cell lines Caco-2 and T84
F Kasuga1, Y Hara-Kudo, N Saito
1Department of Biomedical Food Research, National Institute of Infectious Diseases, Tokyo, Japan.
Abstract:
The effects of low concentrations of deoxynivalenol (DON) on structural and functional characteristics of human colonic adenocarcinoma cell lines Caco-2 and T84 were examined. Scanning electron microscopic (SEM) analysis of the apical surfaces of Caco-2 cells revealed reduction or abnormal formation of brush borders in the presence of 50, 100 and 200 ng/ml of DON. Monolayer integrity of Caco-2 and T84 cells was studied using cells which were cultured on permeable membranes. The transepithelial electrical resistance (TEER) of Caco-2 cells was significantly reduced at 50, 100 and 200 ng/ml of DON, significant increase in lucifer yellow (LY) permeability was also observed in these cells at 100 ng/ml of DON. The TEER of T84 cells was significantly reduced at 100 and 200 ng/ml of DON. LY permeability significantly increased at 200 ng/ml of DON in T84 cells. Enzyme activities in Caco-2 cells were also examined. Alkaline phosphatase activity was reduced from the 6th to 15th day of culture in the presence of 100 or 200 ng/ml of DON, whereas sucrase-isomaltase activity was significantly decreased by adding 50 or 100 ng/ml of DON for 15 or 20 days. Protein content was attenuated only by treatment with 200 ng/ml of DON throughout the experimental period. The results indicate that DON interferes with structural and functional characteristics of differentiation in enterocytes at low doses.
Insights
Low concentrations of deoxynivalenol (DON) disrupt human intestinal cell structure and function. This mycotoxin impairs brush border formation and reduces enzyme activity, impacting enterocyte differentiation.
Area of Science:
- Gastroenterology
- Toxicology
- Cell Biology
Background:
- Deoxynivalenol (DON) is a prevalent mycotoxin found in food.
- Understanding DON's effects on human intestinal cells is crucial for public health.
Purpose of the Study:
- To investigate the impact of low DON concentrations on Caco-2 and T84 human colon cell lines.
- To assess DON's effects on enterocyte structural and functional differentiation.
Main Methods:
- Scanning electron microscopy (SEM) for brush border analysis.
- Transepithelial electrical resistance (TEER) and lucifer yellow (LY) permeability assays for monolayer integrity.
- Enzyme activity assays (alkaline phosphatase, sucrase-isomaltase) and protein content measurement.
Main Results:
- DON reduced brush border formation in Caco-2 cells.
- DON significantly decreased TEER and increased LY permeability in both Caco-2 and T84 cells.
- DON exposure lowered alkaline phosphatase and sucrase-isomaltase activities and protein content in Caco-2 cells.
Conclusions:
- Low-dose DON exposure negatively affects structural and functional differentiation of human enterocytes.
- DON compromises intestinal barrier integrity and cellular enzymatic functions.