Basic fibroblast growth factor-induced low density lipoprotein receptor transcription and surface expression. Signal transduction pathways mediated by the bFGF receptor tyrosine kinase.
Basic fibroblast growth factor (bFGF) has been implicated in the regulation of cell proliferation and cholesterol metabolism. In studies reported herein, we show bFGF increases low density lipoprotein (LDL) binding, uptake, and degradation in arterial smooth muscle cells in a dose-dependent manner. This increase was paralleled by an increase in LDL receptor mRNA steady state levels. To determine if bFGF activated transcription of the LDL receptor gene, we transiently transfected smooth muscle cells with a gene construct consisting of the 5'-upstream promoter region of the DNA from the human LDL receptor gene ligated to a plasmid containing the luciferase gene. We found that bFGF and a protein kinase C (PKC) activator, phorbol 12-myristate 13-acetate, significantly induced luciferase activity driven by the LDL receptor promoter, whereas 25-hydroxycholesterol reduced the luciferase activity in bFGF-stimulated cells. These findings show that bFGF and PKC are inducing LDL receptor gene transcription. We also evaluated potential signal transduction pathways induced by bFGF to establish the mechanism(s) leading to the activation of the LDL receptor gene. Activation of the activity of FGF receptor tyrosine kinase in smooth muscle cells by ligand binding resulted in tyrosine phosphorylation of one of the FGF receptors and a 90-kDa-protein as well as increased tyrosine phosphorylation of phospholipase C-gamma. Parallel observations were made in that increased PKC and protein kinase A activities occurred with bFGF as compared with control cells. Inhibitors of receptor tyrosine kinase and other protein kinases significantly reduced transcription and surface expression of LDL receptor. Finally, several key enzymes that are central to the regulation of LDL-cholesteryl ester metabolism were also studied in bFGF-stimulated cells. An increase in acyl-CoA:cholesterol acyltransferase activity and cholesterol esterification was observed with bFGF stimulation, but there was no effect on the lysosomal or cytoplasmic cholesteryl ester hydrolase activities. Our findings suggest potential signal transduction pathways activated by bFGF which play a role in regulating transcription and surface expression of the LDL receptor.
Hsu
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Department of Biochemistry, Cornell University Medical College, New York, New York 10021.
Nicholson
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Hajjar
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D000818:Animals; D002478:Cells, Cultured; D002784:Cholesterol / Q000378:metabolism; D016222:Fibroblast Growth Factor 2 / Q000494:pharmacology*; D005786:Gene Expression Regulation; D009131:Muscle, Smooth, Vascular; D010750:Phosphoproteins / Q000378:metabolism; D019000:Phosphotyrosine; D012333:RNA, Messenger / Q000235:genetics; D051381:Rats; D020794:Receptor Protein-Tyrosine Kinases / Q000378:metabolism*; D017468:Receptors, Fibroblast Growth Factor / Q000378:metabolism*; D011973:Receptors, LDL / Q000235:genetics / Q000378:metabolism*; D015398:Signal Transduction; D002785:Sterol O-Acyltransferase / Q000378:metabolism; D013755:Tetradecanoylphorbol Acetate / Q000494:pharmacology; D014158:Transcription, Genetic; D014443:Tyrosine / Q000031:analogs & derivatives / Q000378:metabolism
D010750:Phosphoproteins; D012333:RNA, Messenger; D017468:Receptors, Fibroblast Growth Factor; D011973:Receptors, LDL; D016222:Fibroblast Growth Factor 2; D019000:Phosphotyrosine; D014443:Tyrosine; D002784:Cholesterol; D002785:Sterol O-Acyltransferase; D020794:Receptor Protein-Tyrosine Kinases; D013755:Tetradecanoylphorbol Acetate