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Periodontal pathogens promote epithelial-mesenchymal transition in oral squamous carcinoma cells in vitro.

Authors
  • Abdulkareem, A A1, 2
  • Shelton, R M3
  • Landini, G4
  • Cooper, P R2
  • Milward, M R1
  • 1 a Periodontology Unit, Institute of Clinical Sciences, College of Medical and Dental Sciences , The University of Birmingham , Edgbaston, Birmingham , UK.
  • 2 d Oral Biology Unit, School of Dentistry, Institute of Clinical Sciences, College of Medical and Dental Sciences , The University of Birmingham , Edgbaston, Birmingham , UK.
  • 3 b Biomaterials Unit, Institute of Clinical Sciences, College of Medical and Dental Sciences , The University of Birmingham , Edgbaston, Birmingham , UK.
  • 4 c Oral Pathology Unit, Institute of Clinical Sciences, College of Medical and Dental Sciences , The University of Birmingham , Edgbaston, Birmingham , UK.
Type
Published Article
Journal
Cell Adhesion & Migration
Publisher
Landes Bioscience
Publication Date
Sep 05, 2017
Pages
1–11
Identifiers
DOI: 10.1080/19336918.2017.1322253
PMID: 28873015
Source
Medline
Keywords
License
Unknown

Abstract

Epithelial-mesenchymal transition (EMT) is potentially involved in increasing metastasis of oral squamous cell carcinoma (OSCC). Periodontal pathogens are well-known for their ability to induce intense immune responses and here we investigated whether they are involved in inducing EMT. Cultures of OSCC cell line (H400) were treated separately with heat-killed periodontal pathogens F. nucleatum, or P. gingivalis or E. coli LPS for 8 d. EMT-associated features were assayed using sq-PCR and PCR-arrays, for EMT-related markers, and ELISAs for TGF-β1, TNF-α, and EGF. The migratory ability of cells was investigated using scratch and transwell migration assays. E-cadherin and vimentin expression was assessed using immunofluorescence while Snail activation was detected with immunocytochemistry. In addition, the integrity of the cultured epithelial layer was investigated using transepithelial electrical resistance (TEER). PCR data showed significant upregulation after 1, 5, and 8 d in transcription of mesenchymal markers and downregulation of epithelial ones compared with unstimulated controls, which were confirmed by immunofluorescence. Periodontal pathogens also caused a significant increase in level of all cytokines investigated which could be involved in EMT-induction and Snail activation. Exposure of cells to the bacteria increased migration and the rate of wound closure. Downregulation of epithelial markers also resulted in a significant decrease in impedance resistance of cell monolayers to passage of electrical current. These results suggested that EMT was likely induced in OSCC cells in response to stimulation by periodontal pathogens.

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