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J. Biol. Chem., Vol. 275, Issue 37, 28634-28640, September 15, 2000
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From the 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
The Correlation of ATP-binding Cassette 1 mRNA Levels
with Cholesterol Efflux from Various Cell Lines*
,
MCP Hahnemann University, Department of
Biochemistry, Philadelphia, Pennsylvania 19129, the § Joseph
Stokes Jr. Research Institute, The Children's Hospital of
Philadelphia, Philadelphia, Pennsylvania 19104, and ¶ Pfizer Inc.,
Department of Cardiovascular and Metabolic Diseases, Central Research
Division, Groton, Connecticut 06340
/ 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.
*
This work was supported in part by National Institutes of
Health Grants HL22633 and HL07443 and Pfizer Central Research.The costs of publication of this
article were defrayed in part by the
payment of page charges. The article
must therefore be hereby marked
"advertisement" in
accordance with 18 U.S.C. Section
1734 solely to indicate this fact.
To whom correspondence should be addressed: Dept. of
Cardiovascular and Metabolic Diseases, Pfizer Central Research,
118B-206B, Eastern Point Rd., Groton, CT 06340. Tel.: 860-441-4872;
Fax: 860-441-1128; E-mail: omar_l_francone@groton.pfizer.com.
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