Effects of bile acids and steroid/thyroid hormones on the expression of cholesterol 7 alpha-hydroxylase mRNA and the CYP7 gene in HepG2 cells.
The expression of cholesterol 7 alpha-hydroxylase mRNA levels in confluent HepG2 cultures was reduced by tauro- or glyco-conjugates of deoxycholate and chenodeoxycholate, but not by cholate. Ursodeoxycholates, on the other hand, stimulated the mRNA level. The 5'-upstream regions of rat cholesterol 7 alpha-hydroxylase gene (CYP7) were fused to luciferase reporter gene and the constructs, p-3616/Luc, p-224/Luc and p-160/Luc, were transiently transfected into HepG2 cells. Tauro-conjugates of deoxycholate and chenodeoxycholate inhibited the transcriptional activities of the gene constructs in the confluent cells, but not in subconfluent cells. These results reveal that bile acid responsive elements are located in the -160 fragment and also between nt -3616 and -224. Thyroid and steroid hormones stimulated transcriptional activity expressed in the confluent cells and their responsive elements are located upstream of nt -224. It appears that adult phenotypes are responsible for bile acid feedback and hormone response in HepG2 cells.
Crestani
M
M
Department of Biochemistry and Molecular Pathology, Northeastern Ohio University, College of Medicine, Rootstown 44272.
Karam
W G
WG
Chiang
J Y
JY
D000818:Animals; D001647:Bile Acids and Salts / Q000494:pharmacology*; D002478:Cells, Cultured; D002790:Cholesterol 7-alpha-Hydroxylase / Q000235:genetics*; D015971:Gene Expression Regulation, Enzymologic / Q000187:drug effects*; D006801:Humans; D008156:Luciferases / Q000096:biosynthesis / Q000235:genetics; D011401:Promoter Regions, Genetic / Q000235:genetics; D012333:RNA, Messenger / Q000096:biosynthesis*; D051381:Rats; D011993:Recombinant Fusion Proteins / Q000096:biosynthesis; D013256:Steroids / Q000494:pharmacology*; D013963:Thyroid Hormones / Q000494:pharmacology*; D014158:Transcription, Genetic; D014162:Transfection
D001647:Bile Acids and Salts; D012333:RNA, Messenger; D011993:Recombinant Fusion Proteins; D013256:Steroids; D013963:Thyroid Hormones; D008156:Luciferases; D002790:Cholesterol 7-alpha-Hydroxylase