Control of low density lipoprotein receptor gene promoter activity. Ketoconazole inhibits serum lipoprotein but not oxysterol suppression of gene transcription.
We have examined the effects of ketoconazole, a drug which inhibits enzymes involved in cholesterol biosynthesis and metabolism, on the suppressive effects of serum lipoproteins and 25-hydroxycholesterol on low density lipoprotein (LDL) receptor gene promoter activity. A LDL receptor promoter-chloramphenicol acetyltransferase (CAT) fusion gene construct (pLDLR-CAT 6500) was transfected into JEG-3 choriocarcinoma cells, and the transfected cells were cultured in the absence or presence of serum, LDL, or serum and 25-hydroxycholesterol. Serum, LDL, and serum + 25-hydroxycholesterol reduced chloramphenicol acetyltransferase activity in cells transfected with pLDLR-CAT 6500, whereas these treatments had no effect upon enzyme activity in cells transfected with a control construct (pSV2CAT). Ketoconazole (50 microM) overcame the effects of serum and LDL on suppression of pLDLR-CAT 6500 expression, but could not override the combination of serum + 25-hydroxycholesterol. Ketoconazole had no significant effect on expression of pSV2CAT. The drug inhibited cholesterol side chain cleavage enzyme in the cells, but appeared to have no impact on the ability of cells to take up LDL-carried lipids. Our observations are consistent with the idea that serum lipoprotein cholesterol is metabolized to an effector substance which acts to suppress LDL receptor gene transcription. The generation of this effector seems to be sensitive to ketoconazole.
Takagi
K
K
Department of Obstetrics and Gynecology, University of Pennsylvania School of Medicine, Philadelphia 19104.
Alvarez
J G
JG
Favata
M F
MF
Trzaskos
J M
JM
Strauss
J F
JF
D002460:Cell Line; D003001:Cloning, Molecular; D003470:Culture Media; D005786:Gene Expression Regulation; D005796:Genes / Q000187:drug effects*; D006801:Humans; D006888:Hydroxycholesterols / Q000494:pharmacology*; D007654:Ketoconazole / Q000494:pharmacology*; D007700:Kinetics; D008074:Lipoproteins / Q000097:blood*; D011401:Promoter Regions, Genetic / Q000187:drug effects*; D011973:Receptors, LDL / Q000235:genetics*; D013489:Suppression, Genetic; D014158:Transcription, Genetic / Q000187:drug effects*; D014407:Tumor Cells, Cultured / Q000187:drug effects / Q000378:metabolism
D003470:Culture Media; D006888:Hydroxycholesterols; D008074:Lipoproteins; D011973:Receptors, LDL; C007997:25-hydroxycholesterol; D007654:Ketoconazole