Differential regulation of constitutive androstane receptor expression by hepatocyte nuclear factor4alpha isoforms.
Constitutive androstane receptor (CAR; NR1I3) controls the metabolism and elimination of endogenous and exogenous toxic compounds by up-regulating a battery of genes. In this work, we analyzed the expression of human CAR (hCAR) in normal liver during development and in hepatocellular carcinoma (HCC) and investigated the effect of hepatocyte nuclear factor 4alpha isoforms (HNF4alpha1 and HNF4alpha7) on the hCAR gene promoter. By performing functional analysis of hCAR 5'-deletions including mutants, chromatin immunoprecipitation in human hepatocytes, electromobility shift and cotransfection assays, we identified a functional and species-conserved HNF4alpha response element (DR1: ccAGGCCTtTGCCCTga) at nucleotide -144. Both HNF4alpha isoforms bind to this element with similar affinity. However, HNF4alpha1 strongly enhanced hCAR promoter activity whereas HNF4alpha7 was a poor activator and acted as a repressor of HNF4alpha1-mediated transactivation of the hCAR promoter. PGC1alpha stimulated both HNF4alpha1-mediated and HNF4alpha7-mediated hCAR transactivation to the same extent, whereas SRC1 exhibited a marked specificity for HNF4alpha1. Transduction of human hepatocytes by HNF4alpha7-expressing lentivirus confirmed this finding. In addition, we observed a positive correlation between CAR and HNF4alpha1 mRNA levels in human liver samples during development, and an inverse correlation between CAR and HNF4alpha7 mRNA levels in HCC. These observations suggest that HNF4alpha1 positively regulates hCAR expression in normal developing and adult livers, whereas HNF4alpha7 represses hCAR gene expression in HCC.
Pascussi
Jean Marc
JM
INSERM, U632, Montpellier, France.
Robert
Agnes
A
Moreau
Amelie
A
Ramos
Jeanne
J
Bioulac-Sage
Paulette
P
Navarro
Francis
F
Blanc
Pierre
P
Assenat
Eric
E
Maurel
Patrick
P
Vilarem
Marie Jose
MJ
D006528:Carcinoma, Hepatocellular / Q000378:metabolism; D002460:Cell Line; D000090702:Constitutive Androstane Receptor; D005786:Gene Expression Regulation; D051557:Hepatocyte Nuclear Factor 4 / Q000502:physiology*; D006801:Humans; D008099:Liver / Q000254:growth & development / Q000378:metabolism*; D008113:Liver Neoplasms / Q000378:metabolism; D011401:Promoter Regions, Genetic / Q000502:physiology; D020033:Protein Isoforms / Q000502:physiology; D018160:Receptors, Cytoplasmic and Nuclear / Q000096:biosynthesis*; D020218:Response Elements / Q000502:physiology; D014157:Transcription Factors / Q000096:biosynthesis*
D000090702:Constitutive Androstane Receptor; D051557:Hepatocyte Nuclear Factor 4; C000716327:NR1I3 protein, human; D020033:Protein Isoforms; D018160:Receptors, Cytoplasmic and Nuclear; D014157:Transcription Factors