Regulation of squalene epoxidase in HepG2 cells.
Regulation of squalene epoxidase in the cholesterol biosynthetic pathway was studied in a human hepatoma cell line, HepG2 cells. Since the squalene epoxidase activity in cell homogenates was found to be stimulated by the addition of Triton X-100, enzyme activity was determined in the presence of this detergent. Incubation of HepG2 cells for 18 h with L-654,969, a potent competitive inhibitor of 3-hydroxy-3-methylglutaryl coenzyme A (HMG-CoA) reductase, increased squalene epoxidase activity dose-dependently. On the other hand, low density lipoprotein (LDL) and 25-hydroxy-cholesterol decreased the enzyme activity. These results demonstrate that squalene epoxidase is regulated by the concentrations of endogenous and exogenous sterols. The affinity of the enzyme for squalene was not changed by treatment with L-654,969. Cytosolic (S105) fractions, prepared from HepG2 cells treated with or without L-654,969, had no effect on microsomal squalene epoxidase activity of HepG2 cells, in contrast to the stimulating effect of S105 fractions from rat liver homogenate. Mevalonate, LDL, and oxysterol treatment abolished the effect of L-654,969. Simultaneous addition of cycloheximide and actinomycin D also prevented enzyme induction in HepG2 cells. From these results, the change in squalene epoxidase activity is thought to be caused by the change in the amount of enzyme protein. It is further suggested that squalene epoxidase activity is suppressed only by sterols, not by nonsterol derivative(s) of mevalonate, in contrast to the regulation of HMG-CoA reductase.
Hidaka
Y
Y
Central Research Laboratories, Banyu Pharmaceutical Co., Ltd., Tokyo, Japan.
Satoh
T
T
Kamei
T
T
D001667:Binding, Competitive; D006528:Carcinoma, Hepatocellular; D002784:Cholesterol / Q000378:metabolism; D003902:Detergents; D004790:Enzyme Induction; D006801:Humans; D019161:Hydroxymethylglutaryl-CoA Reductase Inhibitors; D007700:Kinetics; D008077:Lipoproteins, LDL / Q000378:metabolism; D008099:Liver / Q000201:enzymology*; D008113:Liver Neoplasms; D008148:Lovastatin / Q000031:analogs & derivatives / Q000494:pharmacology; D008798:Mevalonic Acid / Q000378:metabolism / Q000494:pharmacology; D008862:Microsomes, Liver / Q000201:enzymology; D017830:Octoxynol; D010105:Oxygenases / Q000096:biosynthesis / Q000378:metabolism*; D011092:Polyethylene Glycols / Q000494:pharmacology; D019821:Simvastatin / Q000031:analogs & derivatives*; D050603:Squalene Monooxygenase; D014407:Tumor Cells, Cultured
D003902:Detergents; D019161:Hydroxymethylglutaryl-CoA Reductase Inhibitors; D008077:Lipoproteins, LDL; D011092:Polyethylene Glycols; D017830:Octoxynol; D002784:Cholesterol; C063287:simvastatin acid; D008148:Lovastatin; D019821:Simvastatin; D010105:Oxygenases; D050603:Squalene Monooxygenase; D008798:Mevalonic Acid