Fusobacterium nucleatum Caused DNA Damage and Promoted Cell Proliferation by the Ku70/ p53 Pathway in Oral Cancer Cells

DNA Cell Biol. 2020 Jan;39(1):144-151. doi: 10.1089/dna.2019.5064. Epub 2019 Nov 25.

Abstract

Bacterial infection influences genomic stability and integrity by causing DNA damage, which increases the possibility of tumor initiation and development. We aimed to investigate whether Fusobacterium nucleatum, one of the periodontal pathogens, promoted oral squamous cell carcinoma (OSCC) by causing DNA double-strand break (DSB). Tca8113 tongue squamous cell carcinoma cells were infected with F. nucleatum. The expression of γH2AX was detected by western blots and immunofluorescence. The proliferation and cell cycle alterations were tested by CCK8 and flow cytometry, respectively. The expression levels of Ku70, p53, and p27 were evaluated by quantitative real-time polymerase chain reaction and western blots. A plasmid was used for the overexpression of Ku70 to verify the possible relationship between Ku70 and p53. We confirmed the presence of DSBs in the response to F. nucleatum by detecting the expression of γH2AX. The cell proliferation ability was increased with an accelerated cell cycle while the expression of p27 was decreased. Meanwhile, the expression of Ku70 and wild p53 was downregulated. When Ku70 was overexpressed, the expression of wild p53 in response to F. nucleatum infection was upregulated and cell proliferation was accordingly inhibited. We concluded that F. nucleatum infection promoted the proliferation ability of Tca8113 by causing DNA damage via the Ku70/p53 pathway.

Keywords: DNA damage; Fusobacterium nucleatum; Ku70; OSCC; γH2AX.

MeSH terms

  • Carcinoma, Squamous Cell / genetics
  • Carcinoma, Squamous Cell / microbiology
  • Carcinoma, Squamous Cell / pathology
  • Cell Cycle / genetics
  • Cell Line, Tumor
  • Cell Proliferation / genetics*
  • DNA Breaks, Double-Stranded*
  • Fusobacterium nucleatum / physiology
  • Gene Expression Regulation, Neoplastic
  • Histones / genetics
  • Histones / metabolism
  • Host-Pathogen Interactions
  • Humans
  • Ku Autoantigen / genetics*
  • Ku Autoantigen / metabolism
  • Mouth Neoplasms / genetics
  • Mouth Neoplasms / microbiology
  • Mouth Neoplasms / pathology
  • Signal Transduction / genetics*
  • Tumor Suppressor Protein p53 / genetics*
  • Tumor Suppressor Protein p53 / metabolism

Substances

  • H2AX protein, human
  • Histones
  • Tumor Suppressor Protein p53
  • Ku Autoantigen