Prostanoid– and Interleukin‐1–Induced Primary Genes in Cementoblastic Cells
Copyright policy )Prostanoid– and Interleukin‐1–Induced Primary Genes in Cementoblastic Cells
Background: Cementum is a key component of a functional periodontal organ. However, regenerating lost cementum is difficult and often incomplete. Identifying molecular mediators of cementoblast differentiation and function should lead to better targeted treatment for periodontitis. Prostaglandins increase mineralization of murine cementoblastic OCCM cells and alveolar bone formation, whereas the cytokine interleukin‐1 (IL‐1) inhibits alveolar bone formation. We hypothesized that differentially induced primary genes in OCCM cells may mediate anabolic and catabolic responses. Our objective was to identify primary genes differentially induced by the synthetic prostanoid fluprostenol and IL‐1 in cementoblastic cells.Methods: Confluent OCCM cells were pretreated with the protein synthesis inhibitor cycloheximide followed by fluprostenol or IL‐1 for 1.5 hours. cDNA generated from each group was used for cDNA subtraction hybridization to identify differentially induced genes. Preferential gene induction was verified by Northern blot analysis.Results: Thirteen fluprostenol‐ and seven IL‐1–regulated genes were identified. Among the fluprostenol‐induced genes was mitogen‐activated protein (MAP) kinase phosphatase 1 (MKP1), a negative regulator of MAP kinase signaling. To verify the cDNA subtraction hybridization results, OCCM cells were treated with fluprostenol or prostaglandin F2α (PGF2α), and MKP1 mRNA levels were determined. The 0.001 to 1 μM fluprostenol and 0.01 to 1 μM PGF2α significantly induced MKP1 mRNA levels, which peaked at 1 hour of treatment and returned to baseline at 2 hours.Conclusions: Fluprostenol enhanced, whereas IL‐1 inhibited, OCCM mineralization. Using cDNA subtraction hybridization, we identified primary genes that correlate with the observed anabolic and catabolic responses. These findings further our understanding of cementoblast function and suggest that differentially induced genes may mediate cementum formation and resorption.
- University of California, Los Angeles United States
- Southern California University of Health Sciences United States
- University of California, Berkeley United States
- Touro University California United States
Dental Cementum, DNA, Complementary, Gene Expression Profiling, Nucleic Acid Hybridization, Cell Cycle Proteins, Dual Specificity Phosphatase 1, Blotting, Northern, Dinoprost, Immediate-Early Proteins, Mice, Gene Expression Regulation, Enzyme Induction, Protein Phosphatase 1, Prostaglandins F, Synthetic, Phosphoprotein Phosphatases, Animals, RNA, Messenger, Protein Tyrosine Phosphatases, Cell Line, Transformed, Interleukin-1
Dental Cementum, DNA, Complementary, Gene Expression Profiling, Nucleic Acid Hybridization, Cell Cycle Proteins, Dual Specificity Phosphatase 1, Blotting, Northern, Dinoprost, Immediate-Early Proteins, Mice, Gene Expression Regulation, Enzyme Induction, Protein Phosphatase 1, Prostaglandins F, Synthetic, Phosphoprotein Phosphatases, Animals, RNA, Messenger, Protein Tyrosine Phosphatases, Cell Line, Transformed, Interleukin-1
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