Identification of differentially expressed genes using an annealing control primer system in periodontitis

  • Na, Hee-Sam (Department of Oral Microbiology, School of Dentistry, Pusan National University) ;
  • Kim, Ji-S. (Department of Oral Microbiology, School of Dentistry, Pusan National University) ;
  • Chung, Jin (Department of Oral Microbiology, School of Dentistry, Pusan National University)
  • Received : 2012.08.22
  • Accepted : 2012.09.12
  • Published : 2012.09.30

Abstract

In the gingival tissues of patients with periodontitis, inflammatory responses are mediated by a wide variety of genes. In this study, we screened for differentially expressed genes (DEGs) in periodontitis compared with normal tissue using an annealing control primer (ACP) system. By ACP RT-PCR analysis, we obtained about 160 amplicons, 8 of which were found to be differentially expressed. DEGs in patients with periodontitis were thus successfully and reliably identified by the ACP-based RT PCR technique. The DEGs identified in the screen may also enhance our understanding of the pathogenesis of periodontitis.

Keywords

References

  1. Darveau RP. Periodontitis: a polymicrobial disruption of host homeostasis. Nat Rev Microbiol 2010;8:481-490.
  2. Katz J, Michalek SM. Effect of immune T cells derived from mucosal or systemic tissue on host responses to Porphyromonas gingivalis. Oral Microbiol Immunol 1998; 13:73-80.
  3. Kinane DF, Mooney J, Ebersole JL. Humoral immune response to Actinobacillus actinomycetemcomitans and Porphyromonas gingivalis in periodontal disease. Periodontol 2000 1999;20:289-340.
  4. Hwang IT, Kim YJ, Kim SH, Kwak CI, Gu YY, Chun JY. Annealing control primer system for improving specificity of PCR amplification. Biotechniques 2003;35:1180-1184.
  5. Kim YJ, Kwak CI, Gu YY, Hwang IT, Chun JY. Annealing control primer system for identification of differentially expressed genes on agarose gels. Biotechniques 2004; 36:424-426, 428, 430 passim.
  6. Eskan MA, Benakanakere MR, Rose BG, Zhang P, Zhao J, Stathopoulou P, et al. Interleukin-1beta modulates proinflammatory cytokine production in human epithelial cells. Infect Immun 2008;76:2080-2089.
  7. Gaibani P, Caroli F, Nucci C, Sambri V. Major surface protein complex of Treponema denticola induces the production of tumor necrosis factor alpha, interleukin-1 beta, interleukin-6 and matrix metalloproteinase 9 by primary human peripheral blood monocytes. J Periodontal Res 2010;45:361-366.
  8. Jonsson D, Nebel D, Bratthall G, Nilsson BO. The human periodontal ligament cell: a fibroblast-like cell acting as an immune cell. J Periodontal Res 2010;46:153-157.
  9. Koensgen D, Mustea A, Klaman I, Sun P, Zafrakas M, Lichtenegger W, et al. Expression analysis and RNA localization of PAI-RBP1 (SERBP1) in epithelial ovarian cancer: association with tumor progression. Gynecol Oncol 2007;107:266-273.
  10. Sun W, Guo C, Meng X, Yu Y, Jin Y, Tong D, et al. Differential expression of PAI-RBP1, C1orf142, and COTL1 in non-small cell lung cancer cell lines with differenttumor metastatic potential. J Investig Med 2012; 60:689-694.
  11. Czekay RP, Aertgeerts K, Curriden SA, Loskutoff DJ. Plasminogen activator inhibitor-1 detaches cells from extracellular matrices by inactivating integrins. J Cell Biol 2003;160:781-791.
  12. Czekay RP, Wilkins-Port CE, Higgins SP, Freytag J, Overstreet JM, Klein RM, et al. PAI-1: An Integrator of Cell Signaling and Migration. Int J Cell Biol 2011; 2011:562481.
  13. McMahon GA, Petitclerc E, Stefansson S, Smith E, Wong MK, Westrick RJ, et al. Plasminogen activator inhibitor-1 regulates tumor growth and angiogenesis. J Biol Chem 2001;276:33964-33968.
  14. Kaneko T, Fujii S, Matsumoto A, Goto D, Ishimori N, Watano K, et al. Induction of plasminogen activator inhibitor-1 in endothelial cells by basic fibroblast growth factor and its modulation by fibric acid. Arterioscler Thromb Vasc Biol 2002;22:855-860.
  15. Ma Z, Kwong KY, Tovar JP, Paek D. Cyclic adenosine monophosphate induces plasminogen activator inhibitor-1 expression in human mast cells. Biochem Biophys Res Commun 2010;400:569-574.
  16. Pandey M, Tuncman G, Hotamisligil GS, Samad F. Divergent roles for p55 and p75 TNF-alpha receptors in the induction of plasminogen activator inhibitor-1. Am J Pathol 2003;162:933-941.
  17. Kim SR, Jeong SK, Kim WS, Jeon HJ, Park HJ, Kim MK, et al. The Lipopolysaccharide from Porphyromonas gingivalis induces vascular permeability. International J Oral Biology 2011;36:6.
  18. Dohgu S, Takata F, Matsumoto J, Oda M, Harada E, Watanabe T, et al. Autocrine and paracrine up-regulation of blood-brain barrier function by plasminogen activator inhibitor-1. Microvasc Res 2011;81:103-107.
  19. Buduneli N, Becerik S, Buduneli E, Baylas H, Kinnby B. Gingival status, crevicular fluid tissue-type plasminogen activator, plasminogen activator inhibitor-2 levels in pregnancy versus post-partum. Aust Dent J 2010; 55:292-297.
  20. Kardesler L, Buduneli N, Cetinkalp S, Lappin D, Kinane DF. Gingival crevicular fluid IL-6, tPA, PAI-2, albumin levels following initial periodontal treatment in chronic periodontitis patients with or without type 2 diabetes. Inflamm Res 2010;60:143-151.
  21. Olofsson A, Matsson L, Kinnby B. Plasminogen activating capacity in gingival fluid from deteriorating and stable periodontal pockets. J Periodontal Res 2002;37:60-65.
  22. Kinnby B. The plasminogen activating system in periodontal health and disease. Biol Chem 2002;383: 85-92.
  23. Kucharewicz I, Kowal K, Buczko W, Bodzenta-Lukaszyk A. The plasmin system in airway remodeling. Thromb Res 2003;112:1-7.
  24. Ullbro C, Kinnby B, Lindberg P, Matsson L. Tissue plasminogen activator (t-PA) and placental plasminogen activator inhibitor (PAI-2) in gingival crevicular fluid from patients with Papillon-Lefevre syndrome. J Clin Periodontol 2004;31:708-712.
  25. Deppe H, Hohlweg-Majert B, Holzle F, Kesting MR, Wagenpfeil S, Wolff KD, et al. Content of urokinase-type plasminogen activator (uPA) and its inhibitor PAI-1 in oral mucosa and inflamed periodontal tissue. Quintessence Int 2010;41:165-171.