In nonhepatic cells, cholesterol 7alpha-hydroxylase induces the expression of genes regulating cholesterol biosynthesis, efflux, and homeostasis.
CHO cells expressing the liver-specific gene product cholesterol-7alpha-hydroxylase showed a 6-fold increase in the biosynthesis of [(14)C]cholesterol from [(14)C]acetate, as well as increased enzymatic activities of 3-hydroxy-3-methylglutaryl-coenzyme A (HMG-CoA) reductase and squalene synthase. Cells expressing cholesterol-7alpha-hydroxylase contained less sterol response element-binding protein 1 (SREBP1) precursor, whereas the cellular content of mature SREBP1, as well as the mRNAs of cholesterol biosynthetic genes (HMG-CoA reductase and squalene synthase), were all increased approximately 3-fold. Cells expressing cholesterol-7alpha-hydroxylase displayed greater activities of luciferase reporters containing the SREBP-dependent promoter elements derived from HMG-CoA reductase and farnesyl diphosphate synthase, in spite of accumulating significantly more free and esterified cholesterol and 7alpha-hydroxycholesterol. While cells expressing cholesterol-7alpha-hydroxylase displayed increased SREBP-dependent transcription, sterol-mediated repression of SREBP-dependent transcription by LDL-cholesterol and exogenous oxysterols was similar in both cell types. Cells expressing cholesterol-7alpha-hydroxylase displayed greater rates of secretion of cholesterol as well as increased expression of the ABC1 cassette protein mRNA. Adding 25-hydroxycholesterol to the culture medium of both cell types increased the expression of ABC1 cassette protein mRNA. The combined data suggest that in nonhepatic CHO cells multiple regulatory processes sensitive to cellular sterols act independently to coordinately maintain cellular cholesterol homeostasis.
Spitsen
G M
GM
Mammalian Cell and Molecular Biology Laboratory, San Diego State University, San Diego, CA 92182, USA.
Dueland
S
S
Krisans
S K
SK
Slattery
C J
CJ
Miyake
J H
JH
Davis
R A
RA
D000085:Acetates / Q000378:metabolism; D000818:Animals; D022762:CCAAT-Enhancer-Binding Proteins; D016466:CHO Cells / Q000201:enzymology*; D002478:Cells, Cultured; D002784:Cholesterol / Q000096:biosynthesis* / Q000378:metabolism*; D002790:Cholesterol 7-alpha-Hydroxylase / Q000235:genetics* / Q000378:metabolism; D002788:Cholesterol Esters / Q000378:metabolism; D006224:Cricetinae; D004268:DNA-Binding Proteins / Q000378:metabolism / Q000494:pharmacology; D013186:Farnesyl-Diphosphate Farnesyltransferase / Q000235:genetics / Q000378:metabolism; D015870:Gene Expression; D006706:Homeostasis; D006888:Hydroxycholesterols / Q000494:pharmacology; D006903:Hydroxymethylglutaryl CoA Reductases / Q000235:genetics / Q000378:metabolism; D009687:Nuclear Proteins / Q000378:metabolism / Q000494:pharmacology; D011401:Promoter Regions, Genetic; D012333:RNA, Messenger / Q000378:metabolism; D051381:Rats; D020218:Response Elements; D051780:Sterol Regulatory Element Binding Protein 1; D014157:Transcription Factors; D014158:Transcription, Genetic / Q000187:drug effects
D000085:Acetates; D022762:CCAAT-Enhancer-Binding Proteins; D002788:Cholesterol Esters; D004268:DNA-Binding Proteins; D006888:Hydroxycholesterols; D009687:Nuclear Proteins; D012333:RNA, Messenger; C497389:Srebf1 protein, rat; D051780:Sterol Regulatory Element Binding Protein 1; D014157:Transcription Factors; C007997:25-hydroxycholesterol; D002784:Cholesterol; D006903:Hydroxymethylglutaryl CoA Reductases; D002790:Cholesterol 7-alpha-Hydroxylase; D013186:Farnesyl-Diphosphate Farnesyltransferase