Regulation of the CYP3A4 gene by hydrocortisone and xenobiotics: role of the glucocorticoid and pregnane X receptors.
The molecular mechanisms of regulation of the CYP3A4 gene have been examined in an in vitro reporter gene system, containing -1 kb of the CYP3A4 promoter, in a HepG2 cell line. This system allows for the separate and combined transfection of expression plasmids encoding the human glucocorticoid receptor (hGR) and the human pregnane X receptor (hPXR), and, therefore, the opportunity to assess the role of these receptors in the induction process. Hydrocortisone produces a dose-dependent increase in CYP3A4 activation, a response that is increased in the presence of either receptor. Moreover, transfection of the hPXR decreased the EC(50) for hydrocortisone-dependent induction by a factor of 3.3, a response that was not changed by simultaneous cotransfection of the hGR. In addition, the hydrocortisone dose-response curve falls within the physiological blood level concentration of this steroid, implicating a regulatory role for hydrocortisone in the basal level of CYP3A4 expression. Although the responses to dexamethasone and rifampicin were both increased by both receptors, dexamethasone activation of CYP3A4 was similar for both the hGR and the hPXR, whereas rifampicin-dependent activation favored the hPXR. We conclude that regulation of the CYP3A4 gene is receptor-dependent and that hydrocortisone may function as a regulator of basal expression via the hPXR and the hGR. The implications of this latter conclusion for possible regulatory interactions between hydrocortisone and xenobiotic inducers remain to be clarified.
El-Sankary
W
W
Molecular Toxicology Group, School of Biological Sciences, University of Surrey, Guildford, Surrey, United Kingdom.
Plant
N J
NJ
Gibson
G G
GG
Moore
D J
DJ
D000469:Alkaline Phosphatase / Q000378:metabolism; D000893:Anti-Inflammatory Agents / Q000494:pharmacology*; D002460:Cell Line; D002478:Cells, Cultured; D051544:Cytochrome P-450 CYP3A; D003577:Cytochrome P-450 Enzyme System / Q000235:genetics*; D015971:Gene Expression Regulation, Enzymologic / Q000187:drug effects*; D017930:Genes, Reporter / Q000235:genetics; D006801:Humans; D006854:Hydrocortisone / Q000494:pharmacology*; D008099:Liver / Q000166:cytology / Q000187:drug effects / Q000201:enzymology; D006899:Mixed Function Oxygenases / Q000235:genetics*; D010957:Plasmids / Q000235:genetics; D000077297:Pregnane X Receptor; D018160:Receptors, Cytoplasmic and Nuclear / Q000187:drug effects*; D011965:Receptors, Glucocorticoid / Q000187:drug effects*; D011987:Receptors, Steroid / Q000187:drug effects*; D014162:Transfection / Q000235:genetics; D015262:Xenobiotics / Q000494:pharmacology*
D000893:Anti-Inflammatory Agents; D000077297:Pregnane X Receptor; D018160:Receptors, Cytoplasmic and Nuclear; D011965:Receptors, Glucocorticoid; D011987:Receptors, Steroid; D015262:Xenobiotics; D003577:Cytochrome P-450 Enzyme System; D006899:Mixed Function Oxygenases; C104464:CYP3A protein, human; D051544:Cytochrome P-450 CYP3A; C510163:CYP3A4 protein, human; D000469:Alkaline Phosphatase; D006854:Hydrocortisone