title: Role for PKC alpha and PKC epsilon in down-regulation of CFTR mRNA in a human epithelial liver cell line.
abstract: OBJECTIVE
  In the liver, intrahepatic biliary cells are the sole site of expression of the cystic fibrosis transmembrane conductance regulator, the product of the cystic fibrosis gene. We examined the regulation of cystic fibrosis transmembrane conductance regulator gene expression by protein kinase C in the recently characterized human liver epithelial BC1 cell line which expresses, at early confluence, both biliary (cystic fibrosis transmembrane conductance regulator, cytokeratin 19) and hepatocytic (albumin) specific markers.
  
  
  METHODS
  Expression of the cystic fibrosis transmembrane conductance regulator was examined at the mRNA level by Northern blot, reverse transcription-polymerase chain reaction and nuclear run-on assays and at the protein level by Western blotting. The functionality of this protein was tested by measurement of chloride efflux. Protein kinase C isotype expression and cytosol-to-membrane translocation were analysed by Western blotting.
  
  
  RESULTS
  1) Phorbol ester down-regulated cystic fibrosis transmembrane conductance regulator mRNA expression in a time- and dose-dependent manner through a post-transcriptional mechanism with concomitant inhibition of stimulated chloride efflux. 2) Phorbol ester also activated protein kinase C as indicated by the cytosol-to-membrane translocation of both protein kinase C alpha and epsilon the two major protein kinase C isotypes expressed by BC1 cells. 3) Further, maximal down-regulation of the cystic fibrosis transmembrane conductance regulator mRNA by the phorbol ester was inhibited by H7 and by GF 109203X, two known protein kinase C inhibitors.
  
  
  CONCLUSIONS
  These findings provide the first evidence for phorbol ester-induced down-regulation of cystic fibrosis transmembrane conductance regulator mRNA expression in a human liver epithelial cell line and point to a role for the classical protein kinase C alpha and the novel protein kinase C epsilon in this process.
authors.affiliation: Institut National de la Santé et de la Recherche Médicale U. 402, Faculté de Médecine Saint-Antoine, Paris, France.
authors.forename: S
authors.initials: S
authors.lastname: Kang-Park
authors.forename: N
authors.initials: N
authors.lastname: Dray-Charier
authors.forename: A
authors.initials: A
authors.lastname: Munier
authors.forename: C
authors.initials: C
authors.lastname: Brahimi-Horn
authors.forename: D
authors.initials: D
authors.lastname: Veissiere
authors.forename: J
authors.initials: J
authors.lastname: Picard
authors.forename: J
authors.initials: J
authors.lastname: Capeau
authors.forename: G
authors.initials: G
authors.lastname: Cherqui
authors.forename: O
authors.initials: O
authors.lastname: Lascols
mesh_terms: D002460:Cell Line; D019005:Cystic Fibrosis Transmembrane Conductance Regulator / Q000235:genetics*; D015536:Down-Regulation; D004847:Epithelial Cells / Q000378:metabolism*; D015971:Gene Expression Regulation, Enzymologic / Q000502:physiology; D006207:Half-Life; D006801:Humans; D007527:Isoenzymes / Q000502:physiology*; D008099:Liver / Q000166:cytology / Q000378:metabolism*; D010641:Phenotype; D011493:Protein Kinase C / Q000502:physiology*; D051571:Protein Kinase C-alpha; D051744:Protein Kinase C-epsilon; D012333:RNA, Messenger / Q000378:metabolism*; D013755:Tetradecanoylphorbol Acetate / Q000494:pharmacology; D014407:Tumor Cells, Cultured
chemical_list: C505032:CFTR protein, human; D007527:Isoenzymes; D012333:RNA, Messenger; D019005:Cystic Fibrosis Transmembrane Conductance Regulator; C496628:PRKCA protein, human; C497255:PRKCE protein, human; D011493:Protein Kinase C; D051571:Protein Kinase C-alpha; D051744:Protein Kinase C-epsilon; D013755:Tetradecanoylphorbol Acetate