Skip to main content
SpringerLink
Log in

Transport of the soy isoflavone daidzein and its conjugative metabolites by the carriers SOAT, NTCP, OAT4, and OATP2B1

  • Toxicokinetics and Metabolism
  • Published:
Archives of Toxicology Aims and scope Submit manuscript

Abstract

Soy isoflavones (IF) are phytoestrogens, which interact with estrogen receptors. They are extensively metabolized by glucuronosyltransferases and sulfotransferases, leading to the modulation of their estrogenic activity. It can be assumed that this biotransformation also has a crucial impact on the uptake of IF by active or passive cellular transport mechanisms, but little is known about the transport of IF phase II metabolites into the cell. Therefore, transport assays for phase II metabolites of daidzein (DAI) were carried out using HEK293 cell lines transfected with five human candidate carriers, i.e., organic anion transporter OAT4, sodium-dependent organic anion transporter (SOAT), Na+-taurocholate cotransporting polypeptide (NTCP), apical sodium-dependent bile acid transporter ASBT, and organic anion transporting polypeptide OATP2B1. Cellular uptake was monitored by UHPLC-DAD. DAI monosulfates were transported by the carriers NTCP and SOAT in a sodium-dependent manner, while OAT4-HEK293 cells revealed a partly sodium-dependent transport for these compounds. In contrast, DAI-7,4′-disulfate was only taken up by NTCP-HEK293 cells. DAI-7-glucuronide, but not DAI-4′-glucuronide, was transported exclusively by OATP2B1 in a sodium-independent manner. DAI-7-glucuronide-4′-sulfate, DAI-7-glucoside, and DAI were no substrate of any of the tested carriers. In addition, the inhibitory potency of the DAI metabolites toward estrone-sulfate (E1S) uptake of the above-mentioned carriers was determined. In conclusion, human SOAT, NTCP, OATP2B1, and OAT4 were identified as carriers for the DAI metabolites. Several metabolites were able to inhibit carrier-dependent E1S uptake. These findings might contribute to a better understanding of the bioactivity of IF especially in case of hormone-related cancers.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Log in via an institution

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Fig. 1
Fig. 2
Fig. 3
Fig. 4
Fig. 5

Similar content being viewed by others

Abbreviations

ASBT:

Apical sodium-dependent bile acid transporter

DAI:

Daidzein

DHEAS:

Dehydroepiandrosterone sulfate

DS:

Disulfate

E1S:

Estrone-sulfate

E2S:

17β-Estradiol-3-sulfate

G:

Glucoside

GA:

Glucuronide

GEN:

Genistein

GLY:

Glycitein

IF:

Isoflavones

NTCP:

Na+-taurocholate cotransporting polypeptide

OAT:

Organic anion transporter

OATP:

Organic anion transporting polypeptide

S:

Sulfate

SOAT:

Sodium-dependent organic anion transporter

STS:

Steroid sulfatase

References

  • Al Sarakbi W, Mokbel R, Salhab M, Jiang WG, Reed MJ, Mokbel K (2006) The role of STS and OATP-B mRNA expression in predicting the clinical outcome in human breast cancer. Anticancer Res 26:4985–4990

    CAS  PubMed  Google Scholar 

  • Allred CD, Ju YH, Allred KF, Chang J, Helferich WG (2001) Dietary genistin stimulates growth of estrogen-dependent breast cancer tumors similar to that observed with genistein. Carcinogenesis 22:1667–1673

    Article  CAS  PubMed  Google Scholar 

  • Bolca S, Urpi-Sarda M, Blondeel P, Roche N, Vanhaecke L, Possemiers S, Al-Maharik N, Botting N, De Keukeleire D, Bracke M, Heyerick A, Manach C, Depypere H (2010) Disposition of soy isoflavones in normal human breast tissue. Am J Clin Nutr 91:976–984

    Article  CAS  PubMed  Google Scholar 

  • Brant SR, Yun CH, Donowitz M, Tse CM (1995) Cloning, tissue distribution, and functional analysis of the human Na+/N+ exchanger isoform, NHE3. Am J Physiol 269:C198–C206

    CAS  PubMed  Google Scholar 

  • Cha SH, Sekine T, Kusuhara H, Yu E, Kim JY, Kim DK, Sugiyama Y, Kanai Y, Endou H (2000) Molecular cloning and characterization of multispecific organic anion transporter 4 expressed in the placenta. J Biol Chem 275:4507–4512

    Article  CAS  PubMed  Google Scholar 

  • Craddock AL, Love MW, Daniel RW, Kirby LC, Walters HC, Wong MH, Dawson PA (1998) Expression and transport properties of the human ileal and renal sodium-dependent bile acid transporter. Am J Physiol 274:G157–G169

    CAS  PubMed  Google Scholar 

  • Fietz D, Bakhaus K, Wapelhorst B, Grosser G, Gunther S, Alber J, Doring B, Kliesch S, Weidner W, Galuska CE, Hartmann MF, Wudy SA, Bergmann M, Geyer J (2013) Membrane transporters for sulfated steroids in the human testis—cellular localization, expression pattern and functional analysis. PLoS One 8:e62638

    Article  PubMed Central  CAS  PubMed  Google Scholar 

  • Geisler J (2003) Breast cancer tissue estrogens and their manipulation with aromatase inhibitors and inactivators. J Steroid Biochem Mol Biol 86:245–253

    Article  CAS  PubMed  Google Scholar 

  • Geyer J, Wilke T, Petzinger E (2006) The solute carrier family SLC10: more than a family of bile acid transporters regarding function and phylogenetic relationships. Naunyn Schmiedebergs Arch Pharmacol 372:413–431

    Article  CAS  PubMed  Google Scholar 

  • Geyer J, Doring B, Meerkamp K, Ugele B, Bakhiya N, Fernandes CF, Godoy JR, Glatt H, Petzinger E (2007) Cloning and functional characterization of human sodium-dependent organic anion transporter (SLC10A6). J Biol Chem 282:19728–19741

    Article  CAS  PubMed  Google Scholar 

  • Grosser G, Fietz D, Gunther S, Bakhaus K, Schweigmann H, Ugele B, Brehm R, Petzinger E, Bergmann M, Geyer J (2013) Cloning and functional characterization of the mouse sodium-dependent organic anion transporter Soat (Slc10a6). J Steroid Biochem Mol Biol 138:90–99

    Article  CAS  PubMed  Google Scholar 

  • Grube M, Kock K, Karner S, Reuther S, Ritter CA, Jedlitschky G, Kroemer HK (2006) Modification of OATP2B1-mediated transport by steroid hormones. Mol Pharmacol 70:1735–1741

    Article  CAS  PubMed  Google Scholar 

  • Grube M, Reuther S, Meyer Zu Schwabedissen H, Kock K, Draber K, Ritter CA, Fusch C, Jedlitschky G, Kroemer HK (2007) Organic anion transporting polypeptide 2B1 and breast cancer resistance protein interact in the transepithelial transport of steroid sulfates in human placenta. Drug Metab Dispos 35:30–35

    Article  CAS  PubMed  Google Scholar 

  • Hagenbuch B, Meier PJ (1994) Molecular cloning, chromosomal localization, and functional characterization of a human liver Na+/bile acid cotransporter. J Clin Investig 93:1326–1331

    Article  PubMed Central  CAS  PubMed  Google Scholar 

  • Hagos Y, Stein D, Ugele B, Burckhardt G, Bahn A (2007) Human renal organic anion transporter 4 operates as an asymmetric urate transporter. J Am Soc Nephrol 18:430–439

    Article  CAS  PubMed  Google Scholar 

  • Hilakivi-Clarke L, Andrade JE, Helferich W (2010) Is soy consumption good or bad for the breast? J Nutr 140:2326S–2334S

    Article  PubMed Central  CAS  PubMed  Google Scholar 

  • Hosoda K, Furuta T, Ishii K (2011) Metabolism and disposition of isoflavone conjugated metabolites in humans after ingestion of kinako. Drug Metab Dispos 39:1762–1767

    Article  CAS  PubMed  Google Scholar 

  • Imai Y, Tsukahara S, Asada S, Sugimoto Y (2004) Phytoestrogens/flavonoids reverse breast cancer resistance protein/ABCG2-mediated multidrug resistance. Cancer Res 64:4346–4352

    Article  CAS  PubMed  Google Scholar 

  • Kijkuokool P, Parhar IS, Malaivijitnond S (2006) Genistein enhances N-nitrosomethylurea-induced rat mammary tumorigenesis. Cancer Lett 242:53–59

    Article  CAS  PubMed  Google Scholar 

  • Kinjo J, Tsuchihashi R, Morito K, Hirose T, Aomori T, Nagao T, Okabe H, Nohara T, Masamune Y (2004) Interactions of phytoestrogens with estrogen receptors a and b (III). Estrogenic activities of soy isoflavone aglycones and their metabolites isolated from human urine. Biol Pharm Bull 27:185–188

    Article  CAS  PubMed  Google Scholar 

  • Kobayashi D, Nozawa T, Imai K, Nezu J, Tsuji A, Tamai I (2003) Involvement of human organic anion transporting polypeptide OATP-B (SLC21A9) in pH-dependent transport across intestinal apical membrane. J Pharmacol Exp Ther 306:703–708

    Article  CAS  PubMed  Google Scholar 

  • Koenen A, Kock K, Keiser M, Siegmund W, Kroemer HK, Grube M (2012) Steroid hormones specifically modify the activity of organic anion transporting polypeptides 2852. Eur J Pharm Sci 47:774–780

    Article  CAS  PubMed  Google Scholar 

  • Kramer W, Stengelin S, Baringhaus KH, Enhsen A, Heuer H, Becker W, Corsiero D, Girbig F, Noll R, Weyland C (1999) Substrate specificity of the ileal and the hepatic Na(+)/bile acid cotransporters of the rabbit. I. Transport studies with membrane vesicles and cell lines expressing the cloned transporters. J Lipid Res 40:1604–1617

    CAS  PubMed  Google Scholar 

  • Kullak-Ublick GA, Ismair MG, Stieger B, Landmann L, Huber R, Pizzagalli F, Fattinger K, Meier PJ, Hagenbuch B (2001) Organic anion-transporting polypeptide B (OATP-B) and its functional comparison with three other OATPs of human liver. Gastroenterology 120:525–533

    Article  CAS  PubMed  Google Scholar 

  • Lang K, Wagner C, Haddad G, Burnekova O, Geibel J (2003) Intracellular pH activates membrane-bound Na(+)/H(+) exchanger and vacuolar H(+)-ATPase in human embryonic kidney (HEK) cells. Cell Physiol Biochem 13:257–262

    Article  CAS  PubMed  Google Scholar 

  • Lowry OH, Rosebrough NJ, Farr AL, Randall RJ (1951) Protein measurement with the Folin phenol reagent. J Biol Chem 193:265–275

    CAS  PubMed  Google Scholar 

  • Magee PJ, Rowland I (2012) Soy products in the management of breast cancer. Curr Opin Clin Nutr Metab Care 15:586–591

    Article  CAS  PubMed  Google Scholar 

  • Meier PJ, Stieger B (2002) Bile salt transporters. Annu Rev Physiol 64:635–661

    Article  CAS  PubMed  Google Scholar 

  • Mortensen A, Kulling SE, Schwartz H, Rowland I, Ruefer CE, Rimbach G, Cassidy A, Magee P, Millar J, Hall WL, Birkved FK, Sorensen IK, Sontag G (2009) Analytical and compositional aspects of isoflavones in food and their biological effects. Mol Nutr Food Res 53:S266–S309

    Article  PubMed  Google Scholar 

  • Nishimura M, Naito S (2005) Tissue-specific mRNA expression profiles of human ATP-binding cassette and solute carrier transporter superfamilies. Drug Metab Pharmacokinet 20:452–477

    Article  CAS  PubMed  Google Scholar 

  • Pasqualini JR, Chetrite GS (2005) Recent insight on the control of enzymes involved in estrogen formation and transformation in human breast cancer. J Steroid Biochem Mol Biol 93:221–236

    Article  CAS  PubMed  Google Scholar 

  • Pizzagalli F, Varga Z, Huber RD, Folkers G, Meier PJ, St-Pierre MV (2003) Identification of steroid sulfate transport processes in the human mammary gland. J Clin Endocrinol Metab 88:3902–3912

    Article  CAS  PubMed  Google Scholar 

  • Pugazhendhi D, Watson KA, Mills S, Botting N, Pope GS, Darbre PD (2008) Effect of sulphation on the oestrogen agonist activity of the phytoestrogens genistein and daidzein in MCF-7 human breast cancer cells. J Endocrinol 197:503–515

    Article  PubMed Central  CAS  PubMed  Google Scholar 

  • Reed MJ, Purohit A, Woo LW, Newman SP, Potter BV (2005) Steroid sulfatase: molecular biology, regulation, and inhibition. Endocr Rev 26:171–202

    Article  CAS  PubMed  Google Scholar 

  • Ritchie MR, Cummings JH, Morton MS, Steel CM, Bolton-Smith C, Riches AC (2006) A newly constructed and validated isoflavone database for the assessment of total genistein and daidzein intake. Br J Nutr 95:204–213

    Article  CAS  PubMed  Google Scholar 

  • Roth M, Obaidat A, Hagenbuch B (2012) OATPs, OATs and OCTs: the organic anion and cation transporters of the SLCO and SLC22A gene superfamilies. Br J Pharmacol 165:1260–1287

    Article  PubMed Central  CAS  PubMed  Google Scholar 

  • Rüfer CE, Bub A, Moseneder J, Winterhalter P, Sturtz M, Kulling SE (2008) Pharmacokinetics of the soybean isoflavone daidzein in its aglycone and glucoside form: a randomized, double-blind, crossover study. Am J Clin Nutr 87:1314–1323

    PubMed  Google Scholar 

  • Santner SJ, Feil PD, Santen RJ (1984) In situ estrogen production via the estrone sulfatase pathway in breast tumors: relative importance versus the aromatase pathway. J Clin Endocrinol Metab 59:29–33

    Article  CAS  PubMed  Google Scholar 

  • Schroeder A, Eckhardt U, Stieger B, Tynes R, Schteingart CD, Hofmann AF, Meier PJ, Hagenbuch B (1998) Substrate specificity of the rat liver Na(+)-bile salt cotransporter in Xenopus laevis oocytes and in CHO cells. Am J Physiol 274:G370–G375

    CAS  PubMed  Google Scholar 

  • Schweigmann H, Sanchez-Guijo A, Ugele B, Hartmann K, Hartmann MF, Bergmann M, Pfarrer C, Doring B, Wudy SA, Petzinger E, Geyer J, Grosser G (2014) Transport of the placental estriol precursor 16alpha-hydroxy-dehydroepiandrosterone sulfate (16alpha-OH-DHEAS) by stably transfected OAT4-, SOAT-, and NTCP-HEK293 cells. J Steroid Biochem Mol Biol 143:259–265. doi:10.1016/j.jsbmb.2014.03.013

    Article  CAS  PubMed  Google Scholar 

  • Selcer KW, Kabler H, Sarap J, Xiao Z, Li PK (2002) Inhibition of steryl sulfatase activity in LNCaP human prostate cancer cells. Steroids 67:821–826

    Article  CAS  PubMed  Google Scholar 

  • Sfakianos J, Coward L, Kirk M, Barnes S (1997) Intestinal uptake and biliary excretion of the isoflavone genistein in rats. J Nutr 127:1260–1268

    CAS  PubMed  Google Scholar 

  • Shneider BL, Dawson PA, Christie DM, Hardikar W, Wong MH, Suchy FJ (1995) Cloning and molecular characterization of the ontogeny of a rat ileal sodium-dependent bile acid transporter. J Clin Investig 95:745–754

    Article  PubMed Central  CAS  PubMed  Google Scholar 

  • Soukup ST, Al-Maharik N, Botting N, Kulling SE (2014) Quantification of soy isoflavones and their conjugative metabolites in plasma and urine: an automated and validated UHPLC-MS/MS method for use in large-scale studies. Anal Bioanal Chem 406:6007–6020

    Article  CAS  PubMed  Google Scholar 

  • St-Pierre MV, Hagenbuch B, Ugele B, Meier PJ, Stallmach T (2002) Characterization of an organic anion-transporting polypeptide (OATP-B) in human placenta. J Clin Endocrinol Metab 87:1856–1863

    Article  CAS  PubMed  Google Scholar 

  • Tamai I, Nezu J, Uchino H, Sai Y, Oku A, Shimane M, Tsuji A (2000) Molecular identification and characterization of novel members of the human organic anion transporter (OATP) family. Biochem Biophys Res Commun 273:251–260

    Article  CAS  PubMed  Google Scholar 

  • Trauner M, Boyer JL (2003) Bile salt transporters: molecular characterization, function, and regulation. Physiol Rev 83:633–671

    Article  CAS  PubMed  Google Scholar 

  • Ugele B, Bahn A, Rex-Haffner M (2008) Functional differences in steroid sulfate uptake of organic anion transporter 4 (OAT4) and organic anion transporting polypeptide 2B1 (OATP2B1) in human placenta. J Steroid Biochem Mol Biol 111:1–6

    Article  CAS  PubMed  Google Scholar 

  • Wietrzyk J, Mazurkiewicz M, Madej J, Dzimira S, Grynkiewicz G, Radzikowski C, Opolski A (2004) Genistein alone or combined with cyclophosphamide may stimulate 16/C transplantable mouse mammary cancer growth. Med Sci Monitor 10:BR414–BR419

    CAS  Google Scholar 

  • Wong MH, Oelkers P, Dawson PA (1995) Identification of a mutation in the ileal sodium-dependent bile acid transporter gene that abolishes transport activity. J Biol Chem 270:27228–27234

    Article  CAS  PubMed  Google Scholar 

  • Wong CC, Botting NP, Orfila C, Al-Maharik N, Williamson G (2011) Flavonoid conjugates interact with organic anion transporters (OATs) and attenuate cytotoxicity of adefovir mediated by organic anion transporter 1 (OAT1/SLC22A6). Biochem Pharmacol 81:942–949

    Article  CAS  PubMed  Google Scholar 

  • Zamora-Ros R, Ferrari P, González C, Tjønneland A, Olsen A, Bredsdorff L, Overvad K, Touillaud M, Perquier F, Fagherazzi G, Lukanova A, Tikk K, Aleksandrova K, Boeing H, Trichopoulou A, Trichopoulos D, Dilis V, Masala G, Sieri S, Mattiello A, Tumino R, Ricceri F, Bueno-de-Mesquita HB, Peeters PM, Weiderpass E, Skeie G, Engeset D, Menéndez V, Travier N, Molina-Montes E, Amiano P, Chirlaque M-D, Barricarte A, Wallström P, Sonestedt E, Sund M, Landberg R, Khaw K-T, Wareham N, Travis R, Scalbert A, Ward H, Riboli E, Romieu I (2013) Dietary flavonoid and lignan intake and breast cancer risk according to menopause and hormone receptor status in the European Prospective Investigation into Cancer and Nutrition (EPIC) Study. Breast Cancer Res Treat 139:163–176

    Article  CAS  PubMed  Google Scholar 

  • Zhang Y, Kang H, Li B, Zhang R (2012) Positive effects of soy isoflavone food on survival of breast cancer patients in China. Asian Pac J Cancer Prev 13:479–482

    Article  PubMed  Google Scholar 

Download references

Acknowledgments

We want to thank Klaus Schuh for his skillful technical support. Parts of the work were funded by the German Research Foundation (DFG), Grant KU-1079/10-1. The project is part of the collaborative research project entitled IsoCross “Isoflavones: Cross-species comparison on metabolism, estrogen sensitivity, epigenetics, and carcinogenesis”.

Conflict of interest

The authors declare that they have no conflict of interest.

Author information

Authors and Affiliations

  1. Institute of Pharmacology and Toxicology, JLU Giessen, Giessen, Germany

    Gary Grosser, Barbara Döring & Joachim Geyer

  2. Department of Gynaecology and Obstetrics, University Hospital, LMU Munich, Munich, Germany

    Bernhard Ugele

  3. Department of Safety and Quality of Fruit and Vegetables, Max Rubner-Institut, Haid-und-Neu-Straße 9, 76131, Karlsruhe, Germany

    Sabine E. Kulling & Sebastian T. Soukup

Authors
  1. Gary Grosser
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Barbara Döring
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Bernhard Ugele
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Joachim Geyer
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. Sabine E. Kulling
    View author publications

    You can also search for this author in PubMed Google Scholar

  6. Sebastian T. Soukup
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to Sebastian T. Soukup.

Rights and permissions

Reprints and permissions

About this article

Check for updates. Verify currency and authenticity via CrossMark

Cite this article

Grosser, G., Döring, B., Ugele, B. et al. Transport of the soy isoflavone daidzein and its conjugative metabolites by the carriers SOAT, NTCP, OAT4, and OATP2B1. Arch Toxicol 89, 2253–2263 (2015). https://doi.org/10.1007/s00204-014-1379-3

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s00204-014-1379-3

Keywords

Search

Navigation