[KSCI] Korea Science Citation Index Service

http://dx.doi.org/10.9729/AM.2013.43.1.27

Increase of Grb2 and Ras Proteins and Expression of Growth Factors in LPS Stimulated Odontoblast-like Dental Pulp Cells

Jeong, Soon-Jeong (Department of Oral Histology & Developmental Biology, School of Dentistry, Chosun University)
Jeong, Moon-Jin (Department of Oral Histology & Developmental Biology, School of Dentistry, Chosun University)

Publication Information

Applied Microscopy / v.43, no.1, 2013 , pp. 27-33 More about this Journal

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.

Keywords

Grb2; Ras; Lipopolysaacharide; Odontoblast; Dental pulp cells;

Citations & Related Records

Reference

1	Ferrara N (2005) The role of VEGF in the regulation of physiological and pathological angiogenesis. Mech. Angio. Exp. Supp. 209-231.
2	Foukas L C, Katsoulas H L, Paraskevopoulou N, Metheniti A, Lambropoulou M, and Marmaras V J (1998) 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. J. Biol. Chem. 273, 14813-14818. DOI ScienceOn
3	Frank R M and Veogel J C (1980) Ultrastructure of the human odontoblast process and its mineralisation during dental caries. Caries Res. 14, 367-380. DOI ScienceOn
4	Gospodarowicz D, Plouët J, and Malerstein B (1990) 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, 325-333. DOI
5	Guha M and Mackman N (2001) LPS induction of gene expression in human monocytes. Cell. Signal. 13, 85-94. DOI ScienceOn
6	He H, Li W, Chen S Y, Zhang S, Chen Y T, Hayashida Y, Zhu YT, and Tseng S C (2008) Suppression of activation and induction of apoptosis in RAW264.7 cells by amniotic membrane extract. Invest. Ophthalmol. Vis. Sci. 49, 4468-4475. DOI ScienceOn
7	Jin F, Nathan C F, Radzioch D, and Ding A (1998) Lipopolysacchariderelated stimuli induce expression of the secretory leukocyte protease inhibitor, a macrophage-derived lipopolysaccharide inhibitor. Infect. Immun. 66, 2447-2452.
8	Jontell M, Gunraj M N, Bergenholtz G (1987) Immunocompetent cells in the normal dental pulp. J. Dent. Res. 66, 1149-1153. DOI ScienceOn
9	Staquet M J, Durand S H, Colomb E, Roméas A, Vincent C, Bleicher F, Lebecque S, and Farges J C (2008) Different roles of odontoblasts and fibroblasts in immunity. J. Dent. Res. 87, 256-261. DOI ScienceOn
10	Tammela T, Enholm B, Alitalo K, and Paavonen K (2005) The biology of vascular endothelial growth factors. Cardiovasc. Res. 65, 550-563. DOI ScienceOn
11	Yokose S, Kadokura H, Tajima N, Hasegawa A, Sakagami H, Fujieda K, and Katayama T (2004) Platelet-derived growth factor exerts disparate effects on odontoblast differentiation depending on the dimers in rat dental pulp cells. Cell Tissue Res. 315, 375-384. DOI
12	Zamorano J, Kelly A E, Austrian J, Wang H Y, and Keegan A D (2001) 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, 44-54. DOI ScienceOn
13	Berra E, Pages G, and Pouyssegur J (2000) MAP kinases and hypoxia in the control of VEGF expression. Cancer Metastasis Rev. 19, 139-145. DOI ScienceOn
14	Arvidsson A K, Rupp E, Nanberg E, Downward J, Ronnstrand L, Wennstrom S, Schlessinger J, Heldin C H, and Claesson-Welsh L (1994) Tyr-716 in the platelet-derived growth factor beta-receptor kinase insert is involved in GRB2 binding and Ras activation. Mol. Cell. Biol. 14, 6715-6726.
15	Balic A and Mina M (2010) Characterization of progenitor cells in pulps of murine incisors. J. Dent. Res. 89, 1287-1292. DOI ScienceOn
16	Balic A, Aguila L H, Caimano M J, Francone V P, and Mina M (2010) Characterization of stem and progenitor cells in the dental pulp of erupted and unerupted murine molar. Bone 46, 1639-1651. DOI ScienceOn
17	Butler W T and Ritchie H (1995) The nature and functional significance of dentin extracellular matrix proteins. Int. J. Dev. Biol. 39, 169-179.
18	Bornfeldt K E, Raines E W, Graves L M, Skinner M P, Krebs E G, and Ross R (1995) Platelet-derived growth factor. Distinct signal transduction pathways associated with migration versus proliferation. Ann. N. Y. Acad. Sci. 766, 416-430. DOI
19	Botero T M, Mantellini M G, Song W, Hanks C T, and Nor J E (2003) Effect of lipopolysaccharides on vascular endothelial growth factor expression in mouse pulp cells and macrophages. Eur. J. Oral Sci. 111, 228-234. DOI ScienceOn
20	Botero T M, Shelburne C E, Holland G R, Hanks C T, and Nor J E (2006) TLR4 mediates LPS-induced VEGF expression in odontoblasts. J. Endod. 32, 951-955. DOI ScienceOn
21	Choi B D, Jeong S J, Wang G, Kim H J, Kim B O, Hwang H K, Lim D S, Kim S H, and Jeong M J (2009) Temporal induction of secretory leukocyte protease inhibitor (SLPI) in odontoblasts by lipopolysaccharide and wound infection. J. Endod. 35, 997-1002. DOI ScienceOn
22	Coin P G, Lindroos P M, Bird G S, Osornio-Vargas A R, Roggli V L, and Bonner J C (1996) Lipopolysaccharide up-regulates plateletderived growth factor (PDGF) alpha-receptor expression in rat lung myofibroblasts and enhances response to all PDGF isoforms. J. Immunol. 156, 4797-4806.
23	Costa C A, Vaerten M A, Edwards C A, and Hanks C T (1999) Cytotoxic effects of current dental adhesive systems on immortalized odontoblast cell line MDPC-23. Dent. Mater. 15, 434-441. DOI ScienceOn
24	Dauphinee S M and Karsan A (2006) Lipopolysaccharide signaling in endothelial cells. Lab. Invest. 86, 9-22. DOI ScienceOn
25	Dharmawardana P G, Peruzzi B, Giubellino A, Burke T R, and Bottaro D P (2006) Molecular targeting of growth factor receptor-bound 2 (Grb2) as an anti-cancer strategy. Anticancer Drugs 17, 13-20. DOI ScienceOn
26	Masson K, Liu T, Khan R, Sun J, and Ronnstrand L (2009) A role of Gab2 association in Flt3 ITD mediated Stat5 phosphorylation and cell survival. Br. J. Haematol. 146, 193-202. DOI ScienceOn
27	Lee S H, Choi B D, Jeong S J, Jang H S, Kim B O, Lim D S, Park J C, Wang G L, and Jeong M J (2006) Increased protein of the secretory leukocyte protease inhibitor (SLPI) and the expression of growth factors in NIH3T3 cells by LPS stimulation. Kor. J. Elect. Micros. 36, 165-172.
28	Levin L G, Rudd A, Bletsa A, and Reisner H (1999) Expression of IL-8 by cells of the odontoblast layer in vitro. Eur. J. Oral Sci. 107, 131-137. DOI ScienceOn
29	Liu J, Peng D, Zhu Z, Che D, Tang M, and Li D (1998) The expression of PDGF-B chain mRNA in lung tissue from rats repeatedly infected with mycoplasma pneumoniae. J. Tongji Med. Univ. 18, 216-220. DOI
30	Matsushita K, Motani R, Sakuta T, Nagaoka S, Matsuyama T, Abeyama K, Maruyama H, Takada H, Torii M (1999) Lipopolysaccharide enhances the production of vascular endothelial growth factor by human pulp cells in culture. Infect. Immun. 67, 1633-1639.
31	Saluk-Juszczak J and Wachowicz B (2005) The proinflammatory activity of lipopolysaccharide. Postepy. Biochem. 51, 280-287.
32	Schumann R R, Leong S R, Flaggs G W, Gray P W, Wright S D, Mathison J C, Tobias P S, and Ulevitch R J (1990) Structure and function of lipopolysaccharide binding protein. Science 249, 1429-1431. DOI
33	Silva C J, Soldau K, Christen U, Tobias P S, and Ulevitch R J (2001) 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, 21129-21135. DOI ScienceOn
34	Songyang Z, Shoelson S E, McGlade J, Olivier P, Pawson T, Bustelo X R, Barbacid M, Sabe H, Hanafusa H, Yi T, Ren R, Baltimore D, Ratnofsky S, Feldman R A, and Cantley L C (1994) Specific motifs recognized by the SH2 domains of Csk, 3BP2, fps/fes, GRB-2, HCP, SHC, Syk, and Vav. Mol. Cell. Biol. 14, 2777-2785. DOI ScienceOn