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J Biol Chem, Vol. 273, Issue 24, 14738-14744, June 12, 1998

1,25-Dehydroxyvitamin D₃ Synergism toward Transforming Growth Factor-1-induced AP-1 Transcriptional Activity in Mouse Osteoblastic Cells via Its Nuclear Receptor

Akira Takeshita, Kenichi Imai, Shigeaki Kato, Shigeo Kitano, and Shigemasa Hanazawa

From the Department of Oral Microbiology, Meikai University School of Dentistry, Keyakidai, Sakado City, Saitama 350-02, Japan and the  Institute of Molecular and Cellular Biosciences, University of Tokyo, Bunkyo-Ku, Tokyo 113, Japan

The present study demonstrates 1,25-dehydroxyvitamin D₃ (1-25-(OH)₂D₃) synergism toward transforming growth factor (TGF)-1-induced activation protein-1 (AP-1) activity in mouse osteoblastic MC3T3-E1 cells via the nuclear receptor of the vitamin. 1-25-(OH)₂D₃ synergistically stimulated TGF-1-induced expression of the c-jun gene in the cells but not that of the c-fos gene. We actually showed by a gel mobility shift assay 1-25-(OH)₂D₃ synergism of TGF-1-induced AP-1 binding to the 12-(O-tetradecanoylphorbol-13-acetate response element (TRE). 1-25-(OH)₂D₃ markedly stimulated the transient activity of TGF-1-induced AP-1 in the cells transfected with a TRE-chloramphenicol acetyltransferase (CAT) reporter gene. Also, a synergistic increase in TGF-1-induced CAT activity was observed in the cells cotransfected with an expression vector encoding vitamin D₃ receptor (VDR) and the reporter gene. However, the synergistic CAT activity was inhibited by pretreatment with VDR antisense oligonucleotides. In addition, in a Northern blot assay, we observed 1-25-(OH)₂D₃ synergism of TGF-1-induced expression of the c-jun gene in the cells transfected with the VDR expression vector and also found that the synergistic action was clearly blocked by VDR antisense oligonucleotide pretreatment. The present study strongly suggests a novel positive regulation by 1-25-(OH)₂D₃ of TGF-1-induced AP-1 activity in osteoblasts via "genomic action."

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