Role of insulin-like growth factor-1 signaling in dental fibroblast apoptosis.
BACKGROUND
Studies have shown that periodontal ligament fibroblasts (PDLF) and gingival fibroblasts (GF) respond differently to growth factors in the repair and regeneration of periodontal tissues. The goal of this study was to determine the effects of insulin-like growth factor-1 (IGF-1) signaling on cell apoptosis in PDLF compared to GF.


METHODS
The levels of apoptosis were compared between cultured PDLF and GF by DNA fragmentation assay and trypan blue exclusion assay, either in the presence or absence of IGF-1. The transcript level of upstream signaling molecules, such as IGF binding protein-5 (IGFBP-5), IGF-1 receptor (IGF-1R), and phosphoinositide 3-kinase (PI3K), was studied using reverse transcription-polymerase chain reaction (RT-PCR). Furthermore, the role of IGFBP-5 in IGF-1 signaling was verified by annexin-V staining using flow cytometric analysis.


RESULTS
IGF-1 significantly inhibited the level of DNA fragmentation and decreased trypan blue-positive cells in PDLF compared to GF during serum deprivation. The mRNA expression of IGFBP-5, IGF-1R, and PI3K was constitutively upregulated in PDLF compared to GF. In the presence of exogenous IGFBP-5, the annexin-V-positive cells were significantly decreased in GF after IGF-1 stimulation.


CONCLUSIONS
The present study provides evidence that IGF-1 reduces apoptosis in cultured PDLF compared to GF. Upregulation of IGF-1R and PI3K in PDLF further suggests the activation of IGF signaling in PDLF. In addition, the anti-apoptotic effect of IGF-1 may be facilitated by the upregulation of IGFBP-5 in PDLF.
D019870:1-Phosphatidylinositol 4-Kinase / Q000096:biosynthesis; D017209:Apoptosis / Q000187:drug effects / Q000502:physiology*; D002478:Cells, Cultured; D005347:Fibroblasts / Q000166:cytology; D005881:Gingiva / Q000166:cytology*; D006801:Humans; D018975:Insulin-Like Growth Factor Binding Protein 5 / Q000096:biosynthesis / Q000494:pharmacology / Q000502:physiology; D007334:Insulin-Like Growth Factor I / Q000494:pharmacology / Q000502:physiology*; D010513:Periodontal Ligament / Q000166:cytology*; D012333:RNA, Messenger / Q000096:biosynthesis; D017526:Receptor, IGF Type 1 / Q000096:biosynthesis; D011994:Recombinant Proteins / Q000494:pharmacology; D020133:Reverse Transcriptase Polymerase Chain Reaction; D015398:Signal Transduction; D015854:Up-Regulation
D018975:Insulin-Like Growth Factor Binding Protein 5; D012333:RNA, Messenger; D011994:Recombinant Proteins; D007334:Insulin-Like Growth Factor I; D019870:1-Phosphatidylinositol 4-Kinase; D017526:Receptor, IGF Type 1