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Hoffmann, R. A wiki for the life sciences where authorship matters. Nature Genetics (2008)

Interferon-gamma inhibits 1,25-dihydroxyvitamin D3- stimulated synthesis of bone GLA protein in rat osteosarcoma cells by a pretranslational mechanism.

Interferon-gamma (IFN) is produced by lymphocytes in areas of inflammation and connective tissue destruction. IFN inhibits collagen and DNA synthesis in cultured rat long bones and osteoblastic ROS 17/2.8 cells, suggesting that the periarticular loss of bone that occurs in inflammatory joint diseases may be due to IFN inhibition of bone formation. Since serum levels of bone gla protein ( BGP) have been correlated with the bone formation rate, we studied the effect of IFN on production of this osteoblast-specific protein and steady state BGP messenger RNA (mRNA) levels in ROS 17/2.8 cells. RIA of BGP was done using an antibody raised against rat BGP peptide. BGP synthesis was stimulated with 10(-8) M 1,25-dihydroxyvitamin D3 24 h before and continuously after addition of recombinant rat IFN. IFN (100 U/ml) inhibited BGP secretion 52%, 78%, and 70% in the first, second, and third 24 h periods after IFN treatment, compared to control cells cultured with 1,25-dihydroxyvitamin D3 alone. The ED50 for IFN inhibition of BGP production was 3.3 U/ml (0.29 nM). Pulse labeling with [14C]leucine or [3H]proline during the last 4 h of culture revealed that IFN (3-100 U/ml) did not inhibit total protein secretion into the medium. The percent inhibition of BGP production by IFN was independent of media serum concentration or cell density. IFN (100 U/ml) decreased the steady state level of BGP mRNA as measured by Northern analysis using an oligomeric probe for rat BGP. The decrease in hybridization signal for BGP mRNA was detectable by 1 h after IFN exposure and continued to decline at 6 and 24 h. Treatment with cycloheximide (5 micrograms/ml) blocked the inhibitory effect of IFN on steady state levels of BGP mRNA. These results suggest that IFN may inhibit bone formation by selective inhibition of osteoblast matrix protein production. The mechanism of IFN inhibition of BGP production is, at least in part, pretranslational.[1]


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