Increase of Grb2 and Ras Proteins and Expression of Growth Factors in LPS Stimulated Odontoblast-like Dental Pulp Cells
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Inflammatory cells express the inflammatory cytokines and growth factors induced by lipopolysaccharide (LPS). Odontoblasts are located at the pulp–dentin interface and extend their cell processes far into the dentin where they are the first cells to encounter microorganisms or their products. Therefore, this study examined the expression of some growth factors related to the signal pathway, such as growth factor receptor binding protein 2 (Grb2)-Ras in odontoblast-like dental pulp cells, after a treatment with LPS. After 60 minutes, the mRNA and protein expression levels of Grb2 and Ras were higher in the LPS-treated cells than in the control cells. The level of vascular endothelial growth factor (VEGF) and basic fibroblast growth factor (bFGF) mRNA expression was increased significantly to a level similar to that of Grb2 and Ras at 60 minutes. The platelet-derived growth factor-AA (PDGF-AA) mRNA level was expressed strongly in the odontoblast like dental pulp cells without an association with LPS stimulation. Scanning electron microscopy revealed many extensions of the cytoplasmic processes and the number of processes increased gradually at 30, 60 and 90 minutes after LPS stimulation. From these results VEGF and bFGF expression might be induced through the Grb2-Ras signal transduction pathway in LPS treated odontoblasts.
Abstract
Inflammatory cells express the inflammatory cytokines and growth factors induced by lipopolysaccharide (LPS). Odontoblasts are located at the pulp–dentin interface and extend their cell processes far into the dentin where they are the first cells to encounter microorganisms or their products. Therefore, this study examined the expression of some growth factors related to the signal pathway, such as growth factor receptor binding protein 2 (Grb2)-Ras in odontoblast-like dental pulp cells, after a treatment with LPS. After 60 minutes, the mRNA and protein expression levels of Grb2 and Ras were higher in the LPS-treated cells than in the control cells. The level of vascular endothelial growth factor (VEGF) and basic fibroblast growth factor (bFGF) mRNA expression was increased significantly to a level similar to that of Grb2 and Ras at 60 minutes. The platelet-derived growth factor-AA (PDGF-AA) mRNA level was expressed strongly in the odontoblast like dental pulp cells without an association with LPS stimulation. Scanning electron microscopy revealed many extensions of the cytoplasmic processes and the number of processes increased gradually at 30, 60 and 90 minutes after LPS stimulation. From these results VEGF and bFGF expression might be induced through the Grb2-Ras signal transduction pathway in LPS treated odontoblasts.
참고문헌 (34)
- Arvidsson A K, "Tyr-716 in the platelet-derived growth factor beta-receptor kinase insert is involved in GRB2 binding and Ras activation", Mol. Cell. Biol, 14, pp.6715~6726 (1994)
- Balic A, "Characterization of progenitor cells in pulps of murine incisors", J. Dent. Res, 89, pp.1287~1292 (2010)
- Balic A, "Characterization of stem and progenitor cells in the dental pulp of erupted and unerupted murine molar", Bone, 46, pp.1639~1651 (2010)
- Berra E, "MAP kinases and hypoxia in the control of VEGF expression", Cancer Metastasis Rev, 19, pp.139~145 (2000)
- Bornfeldt K E, "Platelet-derived growth factor. Distinct signal transduction pathways associated with migration versus proliferation", Ann. N. Y. Acad. Sci, 766, pp.416~430 (1995)
- Botero T M, "Effect of lipopolysaccharides on vascular endothelial growth factor expression in mouse pulp cells and macrophages", Eur. J. Oral Sci, 111, pp.228~234 (2003)
- Botero T M, "TLR4 mediates LPS-induced VEGF expression in odontoblasts", J. Endod, 32, pp.951~955 (2006)
- Butler W T, "The nature and functional significance of dentin extracellular matrix proteins", Int. J. Dev. Biol, 39, pp.169~179 (1995)
- Choi B D, "Temporal induction of secretory leukocyte protease inhibitor (SLPI) in odontoblasts by lipopolysaccharide and wound infection", J. Endod, 35, pp.997~1002 (2009)
- Coin P G, "Lipopolysaccharide up-regulates plateletderived growth factor (PDGF) alpha-receptor expression in rat lung myofibroblasts and enhances response to all PDGF isoforms", J. Immunol, 156, pp.4797~4806 (1996)
- Costa C A, "Cytotoxic effects of current dental adhesive systems on immortalized odontoblast cell line MDPC-23", Dent. Mater, 15, pp.434~441 (1999)
- Dauphinee S M, "Lipopolysaccharide signaling in endothelial cells", Lab. Invest, 86, pp.9~22 (2006)
- Dharmawardana P G, "Molecular targeting of growth factor receptor-bound 2 (Grb2) as an anti-cancer strategy", Anticancer Drugs, 17, pp.13~20 (2006)
- Ferrara N, "The role of VEGF in the regulation of physiological and pathological angiogenesis", Mech. Angio. Exp, pp.209~231 (2005)
- Foukas L C, "Phagocytosis of Escherichia coli by insect hemocytes requires both activation of the Ras", mitogen-activated protein kinase signal transduction pathway for attachment and beta3 integrin for internalization, 1998, pp.273, 14813~14818
- Frank R M, "Ultrastructure of the human odontoblast process and its mineralisation during dental caries", Caries Res, 14, pp.367~380 (1980)
- Gospodarowicz D, "Comparison of the ability of basic and acidic fibroblast growth factor to stimulate the proliferation of an established keratinocyte cell line: modulation of their biological effects by heparin, transforming growth factor beta (TGF beta), and epidermal growth factor (EGF)", J. Cell. Physiol, 142, pp.325~333 (1990)
- Guha M, "LPS induction of gene expression in human monocytes", Cell. Signal, 13, pp.85~94 (2001)
- He H, "Suppression of activation and induction of apoptosis in RAW264.7 cells by amniotic membrane extract", Invest. Ophthalmol. Vis. Sci, 49, pp.4468~4475 (2008)
- Jin F, "Lipopolysacchariderelated stimuli induce expression of the secretory leukocyte protease inhibitor, a macrophage-derived lipopolysaccharide inhibitor", Infect. Immun, 66, pp.2447~2452 (1998)
- Jontell M, "mmunocompetent cells in the normal dental pulp", J. Dent. Res, 66, pp.1149~1153 (1987)
- 이상화, "NIH3T3 세포주에서 LPS자극에 의한 분비백혈구단백분해효소억제제(SLPI)의 단백질증가와 성장인자들의 발현", 한국현미경학회지, 36(3), pp.165~172 (2006), 한국현미경학회
- Levin L G, "Expression of IL-8 by cells of the odontoblast layer in vitro", Eur. J. Oral Sci, 107, pp.131~137 (1999)
- Liu J, "The expression of PDGF-B chain mRNA in lung tissue from rats repeatedly infected with mycoplasma pneumoniae", J. Tongji Med. Univ, 18, pp.216~220 (1998)
- Masson K, "A role of Gab2 association in Flt3 ITD mediated Stat5 phosphorylation and cell survival", Br. J. Haematol, 146, pp.193~202 (2009)
- Matsushita K, "Lipopolysaccharide enhances the production of vascular endothelial growth factor by human pulp cells in culture", Infect. Immun, 67, pp.1633~1639 (1999)
- Saluk-Juszczak J, "The proinflammatory activity of lipopolysaccharide", Postepy. Biochem, 51, pp.280~287 (2005)
- Schumann R R, "Structure and function of lipopolysaccharide binding protein", Science, 249, pp.1429~1431 (1990)
- Silva C J, "Lipopolysaccharide is in close proximity to each of the proteins in its membrane receptor complex transfer from CD14 to TLR4 and MD-2", J. Biol. Chem, 276, pp.21129~21135 (2001)
- Songyang Z, "Specific motifs recognized by the SH2 domains of Csk, 3BP2, fps", fes, GRB-2, HCP, SHC, Syk, and Vav, 1994, pp.14, 2777~2785
- Staquet M J, "Different roles of odontoblasts and fibroblasts in immunity", J. Dent. Res, 87, pp.256~261 (2008)
- Tammela T, "The biology of vascular endothelial growth factors", Cardiovasc. Res, 65, pp.550~563 (2005)
- Yokose S, "Platelet-derived growth factor exerts disparate effects on odontoblast differentiation depending on the dimers in rat dental pulp cells", Cell Tissue Res, 315, pp.375~384 (2004)
- Zamorano J, "Costimulation of resting B lymphocytes alters the IL-4-activated IRS2 signaling pathway in a STAT6 independent manner: implications for cell survival and proliferation", Cell Res, 11, pp.44~54 (2001)
