Clones of HepG2 cells were selected that stably express the cDNA for hormone-sensitive lipase (HSL). When cells were cultured in the presence of labelled extracellular oleate, accumulation of labelled fatty acid as cellular triacylglycerol (TAG) was significantly lower in the transfectants compared with the wild-type cells. There was no change in the net rate of phospholipid (PL) synthesis. Culture of cells containing isotopically prelabelled TAG resulted in a greater net loss of TAG from the transfected cells than from the wild-type cells. The excess loss of labelled TAG was primarily due to an increased TAG fatty acid oxidation. Free fatty acid release into the medium was not increased in the transfectants, nor was the very low rate of lipoprotein lipid secretion. Also, there was no increased net trafficking of fatty acids from TAG into PLs. Changes in the 3H:14C ratio of TAG prelabelled with [3H]glycerol and [14C]oleate suggested that none of excess TAG fatty acid released in the transfected cells underwent intracellular re-esterification to TAG prior to oxidation. The results suggest that fatty acids mobilized by HSL are directed immediately into the oxidative pathway and are not available for biosynthetic processes. It appears likely, therefore, that intracellular TAG-derived fatty acids which enter the oxidative pathway exist in a different compartment from those that are directed towards synthesis.
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July 1999
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Research Article|
July 08 1999
Metabolic characteristics of a human hepatoma cell line stably transfected with hormone-sensitive lipase
Richard J. PEASE;
Richard J. PEASE
1
*Department of Biochemistry and Molecular Biology, University College London, Gower Street, London WC1E 6BT, U.K.
1To whom correspondence should be addressed ([email protected]).
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David WIGGINS;
David WIGGINS
†Metabolic Research Laboratory, Oxford Lipid Metabolism Group, Nuffield Department of Clinical Medicine, University of Oxford, Radcliffe Infirmary, Woodstock Road, Oxford OX2 6HE, U.K.
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E. David SAGGERSON;
E. David SAGGERSON
*Department of Biochemistry and Molecular Biology, University College London, Gower Street, London WC1E 6BT, U.K.
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Jeni TREE;
Jeni TREE
†Metabolic Research Laboratory, Oxford Lipid Metabolism Group, Nuffield Department of Clinical Medicine, University of Oxford, Radcliffe Infirmary, Woodstock Road, Oxford OX2 6HE, U.K.
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Geoffrey F. GIBBONS
Geoffrey F. GIBBONS
†Metabolic Research Laboratory, Oxford Lipid Metabolism Group, Nuffield Department of Clinical Medicine, University of Oxford, Radcliffe Infirmary, Woodstock Road, Oxford OX2 6HE, U.K.
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Publisher: Portland Press Ltd
Received:
January 29 1999
Revision Received:
April 13 1999
Accepted:
May 07 1999
Online ISSN: 1470-8728
Print ISSN: 0264-6021
The Biochemical Society, London © 1999
1999
Biochem J (1999) 341 (2): 453–460.
Article history
Received:
January 29 1999
Revision Received:
April 13 1999
Accepted:
May 07 1999
Citation
Richard J. PEASE, David WIGGINS, E. David SAGGERSON, Jeni TREE, Geoffrey F. GIBBONS; Metabolic characteristics of a human hepatoma cell line stably transfected with hormone-sensitive lipase. Biochem J 15 July 1999; 341 (2): 453–460. doi: https://doi.org/10.1042/bj3410453
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