- AMER SOC BIOCHEMISTRY MOLECULAR BIOLOGY INC
We investigated the NF-kappa B transcription factor in osteoclast-like cells. Osteoclast-like cells were differentiated from mouse bone marrow cells in co-culture with mouse calvaria-derived primary osteoblasts in the presence of 1 alpha,25-dihydroxyvitamin D-3 and prostaglandin E(2) in collagen gel-coated dishes. We enriched osteoclastlike cells from the co-cultures by Pronase treatment. When the enriched osteoclast-like cells were treated with phorbol 12-myristate 13-acetate, interleukin-1 (IL-1), calcitonin, or macrophage colony-stimulating factor, only IL-1 activated an NF-kappa B-like factor, which specifically bound to a kappa B motif DNA sequence, as detected by an electrophoretic mobility shift assay. IL-1 also activated NF-kappa B induction in osteoblasts. However, the NF-kappa B-like factor induced by IL-1-stimulated osteoclast-like cells is of smaller molecular size than the factor in osteoblasts, as shown by an electrophoretic mobility shift assay. The NF-kappa B activity of osteoclast-like cells was recognized completely by antibodies against the p50 subunit, and only partially by antibodies against the p65 subunit of NF-kappa B. Antibodies against c-Rel, Rel B, and p52 did not recognize the NF-kappa B-like factor. These results suggest that IL-1 activates an NF-kappa B-like factor in osteoclast-like cells, which contains p50 and p65-related proteins.
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