Effect of selenium on selenoprotein P expression in cultured liver cells.
Selenoprotein P and glutathione peroxidase are selenoproteins that are synthesized by hepatocytes. The production of these selenoproteins by human and rat liver cell lines has been assessed at several levels of selenium supplementation and compared with one another. HepG2 and H4IIE cells were cultured in serum-free medium without selenium supplementation for 48 h; then sodium selenite was added to the medium to give final concentrations of 0, 1, 2.5, 5, or 10 ng selenium/ml medium. After 48 h, selenoprotein P concentration in the medium, cellular glutathione peroxidase activity, and the mRNA levels of the two selenoproteins were determined. Selenium deficiency caused a decrease in selenoprotein mRNA and protein levels. The extent of decrease depended on the cell line examined. In selenium-deprived HepG2 cells, selenoprotein P release decreased to 10% of the release by selenium-replete cells. Under the same conditions, cellular glutathione peroxidase activity decreased to 33%. H4IIE cells showed the opposite results with cellular glutathione peroxidase activity decreasing to 13% and selenoprotein P release decreasing to 40% of selenium-replete cells. The effect of dithiothreitol on secretion of selenoprotein P by H4IIE cells was examined. Selenoprotein P secretion was inhibited by dithiothreitol, suggesting that disulfide bond formation is necessary for secretion of the mature protein.
Hill
K E
KE
Department of Medicine/GI and Clinical Nutrition Research Unit, Vanderbilt University School of Medicine, Nashville, TN 37232, USA. kristina.hill@mcmail.vanderbilt.edu
Chittum
H S
HS
Lyons
P R
PR
Boeglin
M E
ME
Burk
R F
RF
D000818:Animals; D003513:Cycloheximide / Q000494:pharmacology; D005979:Glutathione Peroxidase / Q000378:metabolism*; D006801:Humans; D008099:Liver / Q000378:metabolism*; D011500:Protein Synthesis Inhibitors / Q000494:pharmacology; D011506:Proteins / Q000378:metabolism*; D051381:Rats; D012643:Selenium / Q000378:metabolism*; D051149:Selenoprotein P; D051140:Selenoproteins; D014407:Tumor Cells, Cultured
D011500:Protein Synthesis Inhibitors; D011506:Proteins; D051149:Selenoprotein P; D051140:Selenoproteins; D003513:Cycloheximide; D005979:Glutathione Peroxidase; D012643:Selenium