Progenitor-derived hepatocyte-like (B-13/H) cells metabolise 1'-hydroxyestragole to a genotoxic species via a SULT2B1-dependent mechanism.

Abstract

Rat B-13 progenitor cells are readily converted into functional hepatocyte-like B-13/H cells capable of phase I cytochrome P450-dependent activation of pro-carcinogens and induction of DNA damage. The aim of the present study was to investigate whether the cells are also capable of Phase II sulphotransferase (SULT)-dependent activation of a pro-carcinogen to an ultimate carcinogen. To this end we therefore examined the bioactivation of the model hepatic (hepato- and cholangio-) carcinogen estragole and its proximate SULT1A1-activated genotoxic metabolite 1'-hydroxyestragole. Exposing B-13 or B-13/H cells to estragole (at concentrations up to 1mM) resulted in the production of low levels of 1'-hydroxyestragole, but did not result in detectable DNA damage. Exposing B-13/H cells - but not B-13 cells - to 1'-hydroxyestragole resulted in a dose-dependent increase in DNA damage in comet assays, confirmed by detection of N(2)-(trans-isoestragol-3'-yl)-2'-deoxyguanosine adducts. Genotoxicity was inhibited by general SULT inhibitors, supporting a role for SULTS in the activation of 1-hydroxyestragole in B-13/H cells. However, B-13 and B-13/H cells did not express biologically significant levels of SULT1A1 as determined by qRT-PCR, Western blotting and its associated 7-hydroxycoumarin sulphation activity. B-13 and B-13/H cells expressed - relative to intact rat liver - high levels of SULT2B1 (primarily the b isoform) and SULT4A1 mRNAs and proteins. B-13 and B-13/H cells also expressed the 3'-phosphoadenosine 5'-phosphosulphate synthase 1 required for the generation of activated sulphate cofactor 3'-phosphoadenosine 5'-phosphosulphate. However, only B-13/H cells expressed functional SULT activities towards SULT2B1 substrates DHEA, pregnenolone and 4 methylumbelliferone. Since liver progenitor cells are bi-potential and also form cholangiocytes, we therefore hypothesised that B-13 cells express a cholangiocyte-like SULT profile. To test this hypothesis, the expression of SULTs was examined in liver by RT-PCR and immunohistochemistry. SULT2B1 - but not SULT1A1 - was determined to be expressed in both rat and human cholangiocytes. Since 1'-hydroxyestragole exposure readily produced DNA injury in B-13/H cells, these data suggest that cholangiocarcinomas generated in rats fed estragole may be dependent, in part, on SULT2B1 activation of the 1'-hydroxyestragole metabolite.

• Authors: Philip M Probert, Jeremy M Palmer, Wasma Alhusainy, Aimen O Amer, Ivonne M C M Rietjens, Steven A White, David E Jones, Matthew C Wright
• Journal: Toxicology letters (Toxicol Lett) Vol. 243 Pg. 98-110 (Jan 22 2016) ISSN: 1879-3169 [Electronic] Netherlands
• PMID: 26739637 (Publication Type: Journal Article, Research Support, Non-U.S. Gov't)
• Copyright: Copyright © 2016 Z. Published by Elsevier Ireland Ltd.. All rights reserved.

Chemical References

• Allylbenzene Derivatives
• Anisoles
• Carcinogens
• Hymecromone
• Dehydroepiandrosterone
• 1'-hydroxyestragole
• Pregnenolone
• estragole
• Sulfotransferases
• Arylsulfotransferase
• SULT1A1 protein, human
• SULT2B1 protein, human

Topics

• Allylbenzene Derivatives
• Animals
• Anisoles (administration & dosage, toxicity)
• Arylsulfotransferase (antagonists & inhibitors, genetics, metabolism)
• Carcinogens (administration & dosage, toxicity)
• Cell Line
• Comet Assay
• DNA Damage (drug effects)
• Dehydroepiandrosterone (pharmacology)
• Gene Expression Regulation
• Hepatocytes (drug effects)
• Humans
• Hymecromone (pharmacology)
• Liver (cytology, drug effects, metabolism)
• Pregnenolone (pharmacology)
• Rats
• Sulfotransferases (antagonists & inhibitors, genetics, metabolism)