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The effect of green tea catechins on breast cancer resistance protein activity and intestinal efflux of aflatoxin B1 via breast cancer resistance protein in Caco-2 cells

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Abstract

Aflatoxin B1 (AFB1), a mycotoxin produced by Aspergillus spp., was proved as one of the major causes of human hepatocellular carcinoma (HCC) when chronically consumed. An efflux of AFB1 was reported to be associated with breast cancer resistance protein (BCRP) whose activity could also be modulated by green tea catechins. The purpose of this study was, therefore, to examine the impacts of green tea catechins on BCRP activity in Caco-2 cells by H33342 (bis-benzamide, BCRP substrate) accumulation and AFB1 efflux. Results showed a significant decrease (p < 0.05) of AFB1 in the efflux ratio following the incubation with Ko143, a specific BCRP inhibitor, and sodium fluoride, confirming the association of BCRP in AFB1 efflux transport across the cells. Pre-incubation with green tea and gallate catechins (ECG and EGCG) significantly reduced the efflux ratio of AFB1 (p < 0.05) and significantly increased the intracellular H33342 substrate (p < 0.05) in Caco-2 cells, clearly indicating the inhibitory effects of green tea and gallate catechins on BCRP function. Further study on H33342 accumulation revealed a dose-dependent increment of intracellular H33342 when co-administered with increasing concentrations of AFB1. This result implied a possible role of AFB1 as a BCRP competitive inhibitor. The findings from this study concluded the roles of BCRP as an efflux transporter for AFB1 and could be modulated by the exposure of green tea catechins. Owing to a reduction of its efflux, an inhibitory effect of BCRP when pre-exposed with green tea catechins could be crucial for AFB1 cellular accumulation.

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Abbreviations

ABC:

ATP-binding cassette

AFB1 :

Aflatoxin B1

BCRP:

Breast cancer resistance protein

DMEM:

Dulbecco’s modification of eagle’s medium

DMSO:

Dimethyl sulfoxide

EC:

(−)-Epicatechin

ECG:

(−)-Epicatechin gallate

EGC:

(−)-Epigallocatechin

EGCG:

(−)-Epigallocatechin gallate

H33342:

Hoechst 33342 dye substrate

HPLC:

High performance liquid chromatography

NaF:

Sodium fluoride

P-gp:

P-glycoprotein

Papp:

Apparent permeability coefficients

SPE:

Solid phase extraction

SRB:

Sulforhodamine B

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Acknowledgements

This study was financially supported by a research grant from the National Research Council of Thailand (NRCT) and the Faculty of Pharmaceutical Science, Khon Kaen University, Thailand.

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Authors and Affiliations

  1. Graduate Program in Toxicology, Faculty of Pharmaceutical Sciences, Khon Kaen University, Khon Kaen, 40002, Thailand

    Peeradon Tuntiteerawit & Supatra Porasuphatana

  2. Division of Pharmaceutical Chemistry, Faculty of Pharmaceutical Sciences, Khon Kaen University, Khon Kaen, 40002, Thailand

    Kanokwan Jarukamjorn

  3. Research Group for Pharmaceutical Activities of Natural Products Using Pharmaceutical Biotechnology, Faculty of Pharmaceutical Sciences, Khon Kaen University, Khon Kaen, 40002, Thailand

    Kanokwan Jarukamjorn

  4. Division of Pharmacognosy and Toxicology, Faculty of Pharmaceutical Sciences, Khon Kaen University, Khon Kaen, 40002, Thailand

    Supatra Porasuphatana

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  1. Peeradon Tuntiteerawit
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  2. Kanokwan Jarukamjorn
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  3. Supatra Porasuphatana
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Correspondence to Supatra Porasuphatana.

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Tuntiteerawit, P., Jarukamjorn, K. & Porasuphatana, S. The effect of green tea catechins on breast cancer resistance protein activity and intestinal efflux of aflatoxin B1 via breast cancer resistance protein in Caco-2 cells. Toxicol Res. 36, 293–300 (2020). https://doi.org/10.1007/s43188-019-00032-2

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  • DOI: https://doi.org/10.1007/s43188-019-00032-2

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