Identification by differential display PCR of the selenoprotein thioredoxin reductase as a 1 alpha,25(OH)2-vitamin D3-responsive gene in human osteoblasts--regulation by selenite.
1 alpha, 25(OH)2 vitamin D3 (1,25(OH)2D3) is a potent hormone, stimulating bone cell growth and differentiation. In order to detect novel targets for 1,25(OH)2D3 action, we applied differential display PCR (ddPCR) to human fetal osteoblasts (FOB cells). By ddPCR analysis, we identified the selenoprotein thioredoxin reductase (TRR) as a 1,25(OH)2D3-responsive gene. In FOB cells, the response of TRR mRNA steady state levels to 1,25(OH)2D3 was fast and transient. Maximal stimulation was observed after one hour of 1,25(OH)2D3 treatment, thereafter TRR steady state mRNA levels declined to control levels. This transient response of TRR mRNA was not reflected at the TRR enzyme activity level upon treatment with 1,25(OH)2D3 for up to 48 h. Sodium selenite added to differentiated FOB cells increased TRR enzyme activity 2.6-fold, whereas no selenite effect on TRR mRNA steady state levels was measurable. Our data might provide a link between the induction of a differentiation program by 1,25(OH)2D3 and the expression of the selenium responsive TRR system in human osteoblasts.
Medizinische Poliklinik, Universität Würzburg, Germany.
N
N
Schütze
M
M
Bachthaler
A
A
Lechner
J
J
Köhrle
F
F
Jakob
D015152:Blotting, Northern; D002117:Calcitriol / Q000494:pharmacology*; D004622:Embryo, Mammalian; D015971:Gene Expression Regulation, Enzymologic / Q000187:drug effects*; D006801:Humans; D010006:Osteoblasts / Q000201:enzymology*; D016133:Polymerase Chain Reaction; D012333:RNA, Messenger / Q000032:analysis / Q000378:metabolism; D018038:Sodium Selenite / Q000494:pharmacology*; D013880:Thioredoxin-Disulfide Reductase / Q000235:genetics* / Q000378:metabolism
D012333:RNA, Messenger; D013880:Thioredoxin-Disulfide Reductase; D002117:Calcitriol; D018038:Sodium Selenite