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Antiproliferative activity of the ellagic acid-derived gut microbiota isourolithin A and comparison with its urolithin A isomer: the role of cell metabolism

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Abstract

Purpose

Urolithins, metabolites produced by the gut microbiota from ellagic acid, have been acknowledged with cancer chemopreventive activity. Although urolithin A (Uro-A) has been reported to be the most active one, 10–50 % of humans can also produce the isomer isourolithin A (IsoUro-A). However, no biological activity for IsoUro-A has been reported so far. Herein, we describe for the first time the antiproliferative effect of IsoUro-A, compared to Uro-A, against both human colon cancer (Caco-2) and normal (CCD18-Co) cell lines.

Methods

Cell proliferation was evaluated by MTT and Trypan blue exclusion assays. Cell cycle was analyzed by flow cytometry and apoptosis measured by the Annexin V/PI method. Finally, urolithins metabolism was analyzed by HPLC–DAD-MS/MS.

Results

IsoUro-A inhibited the proliferation of Caco-2 cells in a time- and dose-dependent manner, though it was significantly lower than Uro-A (IC50 = 69.7 ± 4.5 and 49.2 ± 3.8 μM at 48 h, respectively). Both urolithins arrested Caco-2 cell cycle at S and G2/M phases and induced apoptosis at concentrations previously found in human colon tissues. Notably, Caco-2 cells glucuronidated more efficiently IsoUro-A than Uro-A (~50 vs. ~20 % of conversion after 48 h, respectively). Both Uro-A and IsoUro-A glucuronides did not exert antiproliferative effects. In addition, cell growth inhibition was higher in Caco-2 than in normal cells.

Conclusions

IsoUro-A exerts strong antiproliferative activity, which is reduced by the extensive glucuronidation at 9-position in cancer cells. Further studies are needed to elucidate whether the in vitro structure–activity relationship found for Uro-A and IsoUro-A plays any role in humans.

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Abbreviations

ACN:

Acetonitrile

ATCC:

American type culture collection

CRC:

Colorectal cancer

DMSO:

Dimethyl sulfoxide

FBS:

Fetal bovine serum

FITC:

Fluorescein isothiocyanate

Glur:

Glucuronide

HPLC:

High-performance liquid chromatography

IC50 :

Half-maximal inhibitory concentration

IsoUro:

Isourolithin

MEM:

Minimal essential medium

MeOH:

Methanol

MS:

Mass spectrometry

MTT:

3-(4,5-Dimethyl-2-thiazolyl)-2,5-diphenyl-2H-tetrazolium bromide

OD:

Optical density

PBS:

Phosphate-buffered saline

PI:

Propidium iodide

SD:

Standard deviation

UGTs:

UDP-glucuronosyltransferases

Uro:

Urolithins

μM:

Micromolar

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Acknowledgments

This work was funded by the Projects CICYT AGL2011-22447 (MINECO, Spain), 201370E068 (CSIC, Spain) and BACCHUS (FP7-KBBE-2012-6-single stage, European Commission Grant Agreement 312090). AGS and MANS are holders of a ‘Juan de la Cierva’ contract and a FPI grant, respectively, from MINECO.

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Authors and Affiliations

  1. Research Group on Quality, Safety and Bioactivity of Plant Foods, Department of Food Science and Technology, CEBAS-CSIC, Campus de Espinardo, P.O. Box 164, 30100, Murcia, Spain

    Antonio González-Sarrías, María Ángeles Núñez-Sánchez, Rocío García-Villalba, Francisco A. Tomás-Barberán & Juan Carlos Espín

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  1. Antonio González-Sarrías
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  2. María Ángeles Núñez-Sánchez
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  3. Rocío García-Villalba
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  4. Francisco A. Tomás-Barberán
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  5. Juan Carlos Espín
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Correspondence to Antonio González-Sarrías.

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González-Sarrías, A., Núñez-Sánchez, M.Á., García-Villalba, R. et al. Antiproliferative activity of the ellagic acid-derived gut microbiota isourolithin A and comparison with its urolithin A isomer: the role of cell metabolism. Eur J Nutr 56, 831–841 (2017). https://doi.org/10.1007/s00394-015-1131-7

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  • DOI: https://doi.org/10.1007/s00394-015-1131-7

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