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. 2005 Jan;47(1-3):79–88. doi: 10.1007/s10616-005-3753-8

In vitro System for Assessing Dioxin Absorption by Intestinal Epithelial Cells and for Preventing this Absorption by Food Substances

Yayoi Natsume 1,✉, Hideo Satsu 1, Mika Hamada 1, Kazushige Kitamura 2, Naoto Okamoto 2, Makoto Shimizu 1
PMCID: PMC3449813  PMID: 19003047

Abstract

A system for assessing intestinal dioxin absorption was established by applying a Caco-2 cell monolayer and stable dioxin-responsive cell line. The stable dioxin-responsive cell line was established by introducing a plasmid incorporating the human CYP1A1 promoter into human hepatic HepG2 genomic DNA upstream of the luciferase gene. 2,3,7,8-Tetrachlorodibenzodioxin (TCDD) was added to the apical side of differentiated human intestinal epithelial Caco-2 cell monolayers that had been cultured on a semipermeable membrane. The basal medium was taken after an appropriate incubation time and added to the dioxin-responsive cells, the TCDD content then being analyzed by a luciferase assay. The amount of TCDD in the basal medium increased in a dose- and time-dependent manner, the results being sufficiently sensitive and reproducible. The inhibition of TCDD permeability to the Caco-2 cell monolayer by such food substances as chlorophyll, insoluble corn fiber and tea dregs were observed by this in vitro assessment system. The system will therefore be useful to identify food substances having a preventive effect on the intestinal absorption of dioxins.

Keywords: 2,3,7,8-Tetrachlorodibenzo-p-dioxin; Assessment system; Food factor; Freeze-thaw arrangement; Small intestine

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Glossary

AhR

aryl hydrocarbon receptor

3-MC

3-methylcholanthrene

MDR1

multi-drug resistance 1

MRP2

multi-drug resistance- associated protein 2

MXR

mitoxantrone resistance protein

RLU

relative light unit

TBT

tributyltin

TCDD

2,3,7,8-tetrachlorodibenzo-p-dioxin

XRE

xenobiotic responsive element

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