Cell and Tissue Research

, Volume 330, Issue 3, pp 447–460

Expression, localization, and inducibility by bile acids of hepatobiliary transporters in the new polarized rat hepatic cell lines, Can 3−1 and Can 10

  • Doris Cassio
  • Rocio I. R. Macias
  • Brigitte Grosse
  • Jose J. G. Marin
  • Maria J. Monte
Regular Article

Abstract

Sinusoidal and apical transporters are responsible for the uptake and biliary elimination of many compounds by hepatocytes. Few in vitro models are however available for analyzing such functions. The expression and bile-acid inducibility of 13 transporters and two nuclear receptors were investigated in the new rat polarized lines, Can 3−1 and Can 10, and in their unpolarized parent, Fao. The relative abundance of mRNA, the protein level, and their localization were examined by real-time quantitative PCR, Western blotting, immunofluorescence, and confocal microscopy. Compared with rat liver, mRNA levels of Fao cells were: negligible for Bsep/Abcb11; lower for the uptake transporters Ntcp and Oatps; similar for SHP, FXR, and Bcrp/Abcg2; and higher (four–fold to 160-fold) for the efflux pumps Mdr1b/Abcb1b, Mdr2/Abcb4, Mrp1/Abcc1, Mrp2/Abcc2, Mrp3/Abcc3, Abcg5, and Abcg8. This profile was mostly maintained (and improved for Bsep) in Can 10. Some transporters were less well expressed in Can 3−1. In both lines, sinusoidal (Ntcp, Mrp3) and canalicular transporters (Mdr-P-glycoproteins detected with C219 antibody, Mrp2) were localized at their correct poles. Bile-acid effects on polarity and mRNA levels of transporters were analyzed after a 6-day treatment with 50 μM taurocholic, chenodeoxycholic (CDCA), or ursodeoxycholic acid (UDCA). No polarization of Fao cells was induced; Can 10 and Can 3−1 polarity was maintained. CDCA and UDCA induced marked enhancement of the volume of Can 10 bile canaliculi. CDCA upregulated Bsep, Mdr2, SHP, Mdr1b, and Oatp2/1a4 in Can 10 (two- to seven-fold) and in Fao cells. Thus, Can 10 constitutes an attractive polarized model for studying vectorial hepatobiliary transport of endogenous and xenobiotic cholephilic compounds.

Keywords

Hepatobiliary transporters Nuclear receptors Bile acids Hepatocyte polarity Cell models Rat 

Abbreviations

Abc/ABC

ATP-binding cassette

BC

bile canaliculi

Bcrp

breast cancer resistance protein

Bsep/BSEP

bile salt export pump

CDCA

chenodeoxycholic acid

DPPIV

dipeptidylpeptidase IV

FITC

fluorescein isothiocyanate

FXR

farnesoid X-activated receptor

Mdr/MDR

multidrug resistance protein

Mrp/MRP

multidrug resistance-associated protein

Ntcp

Na+-taurocholate co-transporting polypeptide

PBS

phosphate-buffered saline

Oatp/OATP

organic anion transporting polypeptide

PM

plasma membrane

SHP

small heterodimer partner

TCA

taurocholic acid

UDCA

ursodeoxycholic acid

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Copyright information

© Springer-Verlag 2007

Authors and Affiliations

  • Doris Cassio
    • 1
    • 2
    • 4
  • Rocio I. R. Macias
    • 3
  • Brigitte Grosse
    • 1
    • 2
  • Jose J. G. Marin
    • 3
  • Maria J. Monte
    • 3
  1. 1.INSERM, U757University of OrsayOrsayFrance
  2. 2.University of OrsayOrsayFrance
  3. 3.Laboratory of Experimental Hepatology and Drug Targeting (HEVEFARM), CIBERehdUniversity of SalamancaSalamancaSpain
  4. 4.Unité INSERM 757, Bâtiment 443Université Paris-SudOrsay CedexFrance

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