title: Pregnane X Receptor (PXR) expression in colorectal cancer cells restricts irinotecan chemosensitivity through enhanced SN-38 glucuronidation.
abstract: BACKGROUND
  Clinical efficacy of chemotherapy in colorectal cancer is subjected to broad inter-individual variations leading to the inability to predict outcome and toxicity. The topoisomerase I inhibitor irinotecan (CPT-11) is worldwide approved for the treatment of metastatic colorectal cancer and undergoes extensive peripheral and tumoral metabolism. PXR is a xenoreceptor activated by many drugs and environmental compounds regulating the expression of drug metabolism and transport genes in detoxification organs such as liver and gastrointestinal tract. Considering the metabolic pathway of irinotecan and the tissue distribution of Pregnane x Receptor (PXR), we hypothesized that PXR could play a key role in colon cancer cell response to irinotecan.
  
  
  RESULTS
  PXR mRNA expression was quantified by RT-quantitative PCR in a panel of 14 colon tumor samples and their matched normal tissues. PXR expression was modulated in human colorectal cancer cells LS174T, SW480 and SW620 by transfection and siRNA strategies. Cellular response to irinotecan and its active metabolic SN38 was assessed by cell viability assays, HPLC metabolic profiles and mRNA quantification of PXR target genes. We showed that PXR was strongly expressed in colon tumor samples and displayed a great variability of expression. Expression of hPXR in human colorectal cancer cells led to a marked chemoresistance to the active metabolite SN38 correlated with PXR expression level. Metabolic profiles of SN38 showed a strong enhancement of SN38 glucuronidation to the inactive SN38G metabolite in PXR-expressing cells, correlated with an increase of UDPglucuronosyl transferases UGT1A1, UGT1A9 and UGT1A10 mRNAs. Inhibition of PXR expression by lentivirus-mediated shRNA, led to SN38 chemoresistance reversion concomitantly to a decrease of UGT1A1 expression and SN38 glucuronidation. Similarly, PXR mRNA expression levels correlated to UGT1A subfamily expression in human colon tumor biopsies.
  
  
  CONCLUSIONS
  Our results demonstrate that tumoral metabolism of SN38 is affected by PXR and point to potential therapeutic significance of PXR quantification in the prediction of irinotecan response. Furthermore, our observations are pharmacologically relevant since many patients suffering from cancer diseases are often exposed to co-medications, food additives or herbal supplements able to activate PXR. A substantial part of the variability observed among patients might be caused by such interactions.
authors.affiliation: Institut de Génomique Fonctionnelle, Centre National de la Recherche Scientifique (CNRS) UMR5203, Institut National de la Santé et de la Recherche Médicale (INSERM) U661, Université Montpellier 1 et 2, Montpellier F-34094, France.
authors.forename: Caroline
authors.initials: C
authors.lastname: Raynal
authors.forename: Jean-Marc
authors.initials: JM
authors.lastname: Pascussi
authors.forename: Géraldine
authors.initials: G
authors.lastname: Leguelinel
authors.forename: Cyril
authors.initials: C
authors.lastname: Breuker
authors.forename: Jovana
authors.initials: J
authors.lastname: Kantar
authors.forename: Benjamin
authors.initials: B
authors.lastname: Lallemant
authors.forename: Sylvain
authors.initials: S
authors.lastname: Poujol
authors.forename: Caroline
authors.initials: C
authors.lastname: Bonnans
authors.forename: Dominique
authors.initials: D
authors.lastname: Joubert
authors.forename: Frédéric
authors.initials: F
authors.lastname: Hollande
authors.forename: Serge
authors.initials: S
authors.lastname: Lumbroso
authors.forename: Jean-Paul
authors.initials: JP
authors.lastname: Brouillet
authors.forename: Alexandre
authors.initials: A
authors.lastname: Evrard
mesh_terms: D002166:Camptothecin / Q000031:analogs & derivatives* / Q000494:pharmacology; D045744:Cell Line, Tumor; D002470:Cell Survival / Q000187:drug effects; D002851:Chromatography, High Pressure Liquid; D015179:Colorectal Neoplasms / Q000201:enzymology / Q000235:genetics* / Q000473:pathology; D019008:Drug Resistance, Neoplasm / Q000187:drug effects*; D004354:Drug Screening Assays, Antitumor; D015972:Gene Expression Regulation, Neoplastic / Q000187:drug effects; D014453:Glucuronosyltransferase / Q000235:genetics / Q000378:metabolism; D006801:Humans; D000077146:Irinotecan; D000077297:Pregnane X Receptor; D012333:RNA, Messenger / Q000235:genetics / Q000378:metabolism; D034741:RNA, Small Interfering / Q000378:metabolism; D011987:Receptors, Steroid / Q000235:genetics* / Q000378:metabolism; D012293:Rifampin / Q000494:pharmacology; D014162:Transfection; D000099148:UGT1A1 Enzyme
chemical_list: D002166:Camptothecin; D014453:Glucuronosyltransferase; D000077146:Irinotecan; D000077297:Pregnane X Receptor; D012333:RNA, Messenger; D034741:RNA, Small Interfering; D011987:Receptors, Steroid; D012293:Rifampin; D000099148:UGT1A1 Enzyme; C106003:SN38-Glu