Journal of Periodontal and Implant Science

Korean Academy of Periodontology (대한치주과학회)
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Helicobacter pylori inhibited cell proliferation in human periodontal ligament fibroblasts through the Cdc25C/CDK1/cyclinB1 signaling cascade

  • Li, Huanying (Department of Stomatology, Nanfang Hospital, Southern Medical University) ;
  • Liang, Dongsheng (Department of Stomatology, Nanfang Hospital, Southern Medical University) ;
  • Hu, Naiming (Department of Stomatology, Nanfang Hospital, Southern Medical University) ;
  • Dai, Xingzhu (Department of Stomatology, Nanfang Hospital, Southern Medical University) ;
  • He, Jianing (Department of Stomatology, Nanfang Hospital, Southern Medical University) ;
  • Zhuang, Hongmin (Department of Stomatology, Nanfang Hospital, Southern Medical University) ;
  • Zhao, Wanghong (Department of Stomatology, Nanfang Hospital, Southern Medical University)
  • Received : 2019.01.30
  • Accepted : 2019.05.16
  • Published : 2019.06.30
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Abstract

Purpose: Several studies have shown that the oral cavity is a secondary location for Helicobacter pylori colonization and that H. pylori is associated with the severity of periodontitis. This study investigated whether H. pylori had an effect on the periodontium. We established an invasion model of a standard strain of H. pylori in human periodontal ligament fibroblasts (hPDLFs), and evaluated the effects of H. pylori on cell proliferation and cell cycle progression. Methods: Different concentrations of H. pylori were used to infect hPDLFs, with 6 hours of co-culture. The multiplicity of infection in the low- and high-concentration groups was 10:1 and 100:1, respectively. The Cell Counting Kit-8 method and Ki-67 immunofluorescence were used to detect cell proliferation. Flow cytometry, quantitative real-time polymerase chain reaction, and western blots were used to detect cell cycle progression. In the high-concentration group, the invasion of H. pylori was observed by transmission electron microscopy. Results: It was found that H. pylori invaded the fibroblasts, with cytoplasmic localization. Analyses of cell proliferation and flow cytometry showed that H. pylori inhibited the proliferation of periodontal fibroblasts by causing G2 phase arrest. The inhibition of proliferation and G2 phase arrest were more obvious in the high-concentration group. In the low-concentration group, the G2 phase regulatory factors cyclin dependent kinase 1 (CDK1) and cell division cycle 25C (Cdc25C) were upregulated, while cyclin B1 was inhibited. However, in the high-concentration group, cyclin B1 was upregulated and CDK1 was inhibited. Furthermore, the deactivated states of tyrosine phosphorylation of CDK1 (CDK1-Y15) and serine phosphorylation of Cdc25C (Cdc25C-S216) were upregulated after H. pylori infection. Conclusions: In our model, H. pylori inhibited the proliferation of hPDLFs and exerted an invasive effect, causing G2 phase arrest via the Cdc25C/CDK1/cyclin B1 signaling cascade. Its inhibitory effect on proliferation was stronger in the high-concentration group.

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References

  1. Lina TT, Alzahrani S, Gonzalez J, Pinchuk IV, Beswick EJ, Reyes VE. Immune evasion strategies used by Helicobacter pylori. World J Gastroenterol 2014;20:12753-66. https://doi.org/10.3748/wjg.v20.i36.12753
  2. Yee JK. Helicobacter pylori colonization of the oral cavity: a milestone discovery. World J Gastroenterol 2016;22:641-8. https://doi.org/10.3748/wjg.v22.i2.641
  3. Krajden S, Fuksa M, Anderson J, Kempston J, Boccia A, Petrea C, et al. Examination of human stomach biopsies, saliva, and dental plaque for Campylobacter pylori. J Clin Microbiol 1989;27:1397-8. https://doi.org/10.1128/JCM.27.6.1397-1398.1989
  4. Dye BA, Kruszon-Moran D, McQuillan G. The relationship between periodontal disease attributes and Helicobacter pylori infection among adults in the United States. Am J Public Health 2002;92:1809-15. https://doi.org/10.2105/AJPH.92.11.1809
  5. Souto R, Colombo AP. Detection of Helicobacter pylori by polymerase chain reaction in the subgingival biofilm and saliva of non-dyspeptic periodontal patients. J Periodontol 2008;79:97-103. https://doi.org/10.1902/jop.2008.070241
  6. Al Asqah M, Al Hamoudi N, Anil S, Al Jebreen A, Al-Hamoudi WK. Is the presence of Helicobacter pylori in dental plaque of patients with chronic periodontitis a risk factor for gastric infection? Can J Gastroenterol 2009;23:177-9. https://doi.org/10.1155/2009/950527
  7. Nonnenmacher C, Dalpke A, Rochon J, Flores-de-Jacoby L, Mutters R, Heeg K. Real-time polymerase chain reaction for detection and quantification of bacteria in periodontal patients. J Periodontol 2005;76:1542-9. https://doi.org/10.1902/jop.2005.76.9.1542
  8. Tatakis DN, Kumar PS. Etiology and pathogenesis of periodontal diseases. Dent Clin North Am 2005;49:491-516. https://doi.org/10.1016/j.cden.2005.03.001
  9. Mysak J, Podzimek S, Sommerova P, Lyuya-Mi Y, Bartova J, Janatova T, et al. Porphyromonas gingivalis: major periodontopathic pathogen overview. J Immunol Res 2014;2014:476068. https://doi.org/10.1155/2014/476068
  10. Socransky SS, Haffajee AD. Dental biofilms difficult therapeutic targets. Periodontol 2000 2002;28:12-55. https://doi.org/10.1034/j.1600-0757.2002.280102.x
  11. Andersen RN, Ganeshkumar N, Kolenbrander PE. Helicobacter pylori adheres selectively to Fusobacterium spp. Oral Microbiol Immunol 1998;13:51-4. https://doi.org/10.1111/j.1399-302X.1998.tb00751.x
  12. Socransky SS, Haffajee AD, Cugini MA, Smith C, Kent RL Jr. Microbial complexes in subgingival plaque. J Clin Periodontol 1998;25:134-44. https://doi.org/10.1111/j.1600-051X.1998.tb02419.x
  13. Okuda K, Kimizuka R, Katakura A, Nakagawa T, Ishihara K. Ecological and immunopathological implications of oral bacteria in Helicobacter pylori-infected disease. J Periodontol 2003;74:123-8. https://doi.org/10.1902/jop.2003.74.1.123
  14. Hu Z, Zhang Y, Li Z, Yu Y, Kang W, Han Y, et al. Effect of Helicobacter pylori infection on chronic periodontitis by the change of microecology and inflammation. Oncotarget 2016;7:66700-12. https://doi.org/10.18632/oncotarget.11449
  15. Al Sayed A, Anand PS, Kamath KP, Patil S, Preethanath RS, Anil S. Oral cavity as an extragastric reservoir of Helicobacter pylori. ISRN Gastroenterol 2014;2014:261369. https://doi.org/10.1155/2014/261369
  16. Zu L. Reviews of research on the relationship between oral Helicobacter pylori and H. pylori infection. Infect Int 2016;5:5-10. https://doi.org/10.1515/ii-2017-0121
  17. Salehi MR, Shah Aboei M, Naghsh N, Hajisadeghi S, Ajami E. A comparison in prevalence of Helicobacter pylori in the gingival crevicular fluid from subjects with periodontitis and healthy individuals using polymerase chain reaction. J Dent Res Dent Clin Dent Prospect 2013;7:238-43.
  18. Chu YT, Wang YH, Wu JJ, Lei HY. Invasion and multiplication of Helicobacter pylori in gastric epithelial cells and implications for antibiotic resistance. Infect Immun 2010;78:4157-65. https://doi.org/10.1128/IAI.00524-10
  19. Evans DG, Evans DJ Jr, Graham DY. Adherence and internalization of Helicobacter pylori by HEp-2 cells. Gastroenterology 1992;102:1557-67. https://doi.org/10.1016/0016-5085(92)91714-F
  20. Dubois A, Boren T. Helicobacter pylori is invasive and it may be a facultative intracellular organism. Cell Microbiol 2007;9:1108-16. https://doi.org/10.1111/j.1462-5822.2007.00921.x
  21. Huang Y, Wang QL, Cheng DD, Xu WT, Lu NH. Adhesion and invasion of gastric mucosa epithelial cells by Helicobacter pylori. Front Cell Infect Microbiol 2016;6:159.
  22. Petersen AM, Krogfelt KA. Helicobacter pylori: an invading microorganism? A review. FEMS Immunol Med Microbiol 2003;36:117-26. https://doi.org/10.1016/S0928-8244(03)00020-8
  23. Necchi V, Candusso ME, Tava F, Luinetti O, Ventura U, Fiocca R, et al. Intracellular, intercellular, and stromal invasion of gastric mucosa, preneoplastic lesions, and cancer by Helicobacter pylori. Gastroenterology 2007;132:1009-23. https://doi.org/10.1053/j.gastro.2007.01.049
  24. Amieva MR, Salama NR, Tompkins LS, Falkow S. Helicobacter pylori enter and survive within multivesicular vacuoles of epithelial cells. Cell Microbiol 2002;4:677-90. https://doi.org/10.1046/j.1462-5822.2002.00222.x
  25. Bode G, Mauch F, Malfertheiner P. The coccoid forms of Helicobacter pylori. Criteria for their viability. Epidemiol Infect 1993;111:483-90. https://doi.org/10.1017/S0950268800057216
  26. Cuddihy AR, O'Connell MJ. Cell-cycle responses to DNA damage in G2. Int Rev Cytol 2003;222:99-140. https://doi.org/10.1016/S0074-7696(02)22013-6
  27. Strauss B, Harrison A, Coelho PA, Yata K, Zernicka-Goetz M, Pines J. Cyclin B1 is essential for mitosis in mouse embryos, and its nuclear export sets the time for mitosis. J Cell Biol 2018;217:179-93. https://doi.org/10.1083/jcb.201612147
  28. Santos SD, Wollman R, Meyer T, Ferrell JE Jr. Spatial positive feedback at the onset of mitosis. Cell 2012;149:1500-13. https://doi.org/10.1016/j.cell.2012.05.028
  29. Schmidt M, Rohe A, Platzer C, Najjar A, Erdmann F, Sippl W. Regulation of G2/M transition by inhibition of WEE1 and PKMYT1 kinases. Molecules 2017;22:E2045. https://doi.org/10.3390/molecules22122045
  30. Sur S, Agrawal DK. Phosphatases and kinases regulating CDC25 activity in the cell cycle: clinical implications of CDC25 overexpression and potential treatment strategies. Mol Cell Biochem 2016;416:33-46. https://doi.org/10.1007/s11010-016-2693-2

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