Applied Microscopy

  • 제43권1호
  • /
  • Pages.27-33
  • /
  • 2013
  • /
  • 2287-5123(pISSN)
  • /
  • 2287-4445(eISSN)
한국현미경학회 (Korean Society of Microscopy)
권호 표지
DOI QR코드

DOI QR Code

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)
  • 투고 : 2013.02.27
  • 심사 : 2013.03.13
  • 발행 : 2013.03.30
PDF 다운로드
⟨ 이전 논문 다음 논문 ⟩

초록

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.

키워드

참고문헌

  1. 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.
  2. Balic A and Mina M (2010) Characterization of progenitor cells in pulps of murine incisors. J. Dent. Res. 89, 1287-1292. https://doi.org/10.1177/0022034510375828
  3. 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. https://doi.org/10.1016/j.bone.2010.02.019
  4. Berra E, Pages G, and Pouyssegur J (2000) MAP kinases and hypoxia in the control of VEGF expression. Cancer Metastasis Rev. 19, 139-145. https://doi.org/10.1023/A:1026506011458
  5. 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. https://doi.org/10.1111/j.1749-6632.1995.tb26691.x
  6. 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. https://doi.org/10.1034/j.1600-0722.2003.00041.x
  7. 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. https://doi.org/10.1016/j.joen.2006.03.018
  8. Butler W T and Ritchie H (1995) The nature and functional significance of dentin extracellular matrix proteins. Int. J. Dev. Biol. 39, 169-179.
  9. 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. https://doi.org/10.1016/j.joen.2009.04.008
  10. 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.
  11. 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. https://doi.org/10.1016/S0109-5641(99)00071-8
  12. Dauphinee S M and Karsan A (2006) Lipopolysaccharide signaling in endothelial cells. Lab. Invest. 86, 9-22. https://doi.org/10.1038/labinvest.3700366
  13. 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. https://doi.org/10.1097/01.cad.0000185180.72604.ac
  14. Ferrara N (2005) The role of VEGF in the regulation of physiological and pathological angiogenesis. Mech. Angio. Exp. Supp. 209-231.
  15. 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. https://doi.org/10.1074/jbc.273.24.14813
  16. 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. https://doi.org/10.1159/000260479
  17. 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. https://doi.org/10.1002/jcp.1041420215
  18. Guha M and Mackman N (2001) LPS induction of gene expression in human monocytes. Cell. Signal. 13, 85-94. https://doi.org/10.1016/S0898-6568(00)00149-2
  19. 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. https://doi.org/10.1167/iovs.08-1781
  20. 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.
  21. Jontell M, Gunraj M N, Bergenholtz G (1987) Immunocompetent cells in the normal dental pulp. J. Dent. Res. 66, 1149-1153. https://doi.org/10.1177/00220345870660061101
  22. 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.
  23. 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. https://doi.org/10.1046/j.0909-8836.1999.eos107209.x
  24. 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. https://doi.org/10.1007/BF02886476
  25. 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. https://doi.org/10.1111/j.1365-2141.2009.07725.x
  26. 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.
  27. Saluk-Juszczak J and Wachowicz B (2005) The proinflammatory activity of lipopolysaccharide. Postepy. Biochem. 51, 280-287.
  28. 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. https://doi.org/10.1126/science.2402637
  29. 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. https://doi.org/10.1074/jbc.M009164200
  30. 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. https://doi.org/10.1128/MCB.14.4.2777
  31. 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. https://doi.org/10.1177/154405910808700304
  32. Tammela T, Enholm B, Alitalo K, and Paavonen K (2005) The biology of vascular endothelial growth factors. Cardiovasc. Res. 65, 550-563. https://doi.org/10.1016/j.cardiores.2004.12.002
  33. 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. https://doi.org/10.1007/s00441-003-0839-5
  34. 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. https://doi.org/10.1038/sj.cr.7290065