Description
The transcriptional regulation of the rat cholesterol 7 alpha-hydroxylase gene (CYP7) by hormones and signal transduction pathways was studied by transient transfection assay of the promoter activity. HepG2 cells were transfected with deletion mutants of the CYP7 upstream region linked to the luciferase reporter gene. The transcription of CYP7/luciferase chimeric genes was higher in confluent than in subconfluent cultures of HepG2 cells. Glucocorticoid receptors, in the presence of dexamethasone, up-regulated the CYP7 gene through two regions located between -3262 and -2803, and between -344 and -222, respectively. Thyroid hormones did not have any effect on the promoter activity. Insulin inhibited the promoter activity through sequences located between -344 and -222, and abolished the stimulation by dexamethasone. Hence, the insulin effect was dominant over that of glucocorticoids. Treatment of transfected HepG2 cells with phorbol
Subject
Animals; Rats; Gene Expression Regulation; Cell Count; Transfection; Base Sequence; Molecular Sequence Data; Phorbol Esters/pharmacology; Cholesterol 7-alpha-Hydroxylase/*genetics; Luciferases/genetics; Cyclic AMP/pharmacology; Hormones/*pharmacology; Second Messenger Systems/physiology; Cultured; Genetic; Tumor Cells; Cloning; Molecular; *Promoter Regions; *Transcription; Enzymologic/*physiology