The correlation of ATP-binding cassette 1 mRNA levels with cholesterol efflux from various cell lines.
Studies show that lipid-free apoA-I stimulates release of cholesterol and phospholipid from fibroblasts and macrophages. ATP-binding cassette 1 (ABC1) is implicated in this release and has been identified as the genetic defect in Tangier disease, evidence that ABC1 is critical to the biogenesis of high density lipoprotein. We quantified levels of ABC1 mRNA, protein, and cholesterol efflux from J774 mouse macrophages +/- exposure to a cAMP analog. Up-regulating ABC1 mRNA correlated to increased cholesterol efflux in a dose- and time-dependent manner. mRNA levels rose after 15 min of exposure while protein levels rose after 1 h, with increased efflux 2-4 h post-treatment. In contrast to cells from wild-type mice, peritoneal macrophages from the Abc1 -/- mouse showed a lower level of basal efflux and no increase with cAMP treatment. The stimulation of efflux exhibits specificity for apoA-I, high density lipoprotein, and other apolipoproteins as cholesterol acceptors, but not for small unilamellar vesicles, bile acid micelles, or cyclodextrin. We have studied a number of cell types and found that while other cell lines express ABC1 constitutively, only J774 and elicited mouse macrophages show a substantial increase of mRNA and efflux with cAMP treatment. ApoA-I-stimulated efflux was detected from the majority of cell lines examined, independent of treatment.
Bortnick
A E
AE
MCP Hahnemann University, Department of Biochemistry, Philadelphia, Pennsylvania 19129, USA.
Rothblat
G H
GH
Stoudt
G
G
Hoppe
K L
KL
Royer
L J
LJ
McNeish
J
J
Francone
O L
OL
D064286:ATP Binding Cassette Transporter 1; D018528:ATP-Binding Cassette Transporters / Q000235:genetics*; D000818:Animals; D002460:Cell Line; D002784:Cholesterol / Q000378:metabolism*; D000242:Cyclic AMP / Q000031:analogs & derivatives / Q000494:pharmacology; D006023:Glycoproteins / Q000235:genetics*; D006801:Humans; D008075:Lipoproteins, HDL / Q000096:biosynthesis; D008264:Macrophages / Q000378:metabolism; D051379:Mice; D012333:RNA, Messenger / Q000032:analysis*; D013873:Thionucleotides / Q000494:pharmacology
C577836:ABCA1 protein, human; D064286:ATP Binding Cassette Transporter 1; D018528:ATP-Binding Cassette Transporters; D006023:Glycoproteins; D008075:Lipoproteins, HDL; D012333:RNA, Messenger; D013873:Thionucleotides; C015929:8-((4-chlorophenyl)thio)cyclic-3',5'-AMP; D002784:Cholesterol; D000242:Cyclic AMP