MicroRNAs Regulate Cytokine Responses in Gingival Epithelial Cells

Steven C Y Chen, Christos Constantinides, Moritz Kebschull, Panos N Papapanou

Research output: Contribution to journalArticlepeer-review

14 Citations (Scopus)

Abstract

MicroRNAs (miRNAs) have been established as key regulators of various biological processes with possible involvement in the pathobiology of periodontal disease. Expanding our earlier observations of substantial differential expression of specific miRNAs between clinically healthy and periodontitis-affected gingival tissues, we used miRNA inhibitors (sponges) in loss-of-function experiments to investigate the involvement of specific miRNAs in the response of pocket epithelium-derived, telomerase-immortalized human gingival keratinocytes (TIGKs) to microbial infection. We constructed stable knockdown (KD) cell lines for five epithelium-expressed miRNAs (miR-126, miR-141, miR-155, miR-210, and miR-1246) and assessed their response to infection with periodontal pathogens using microarray analysis, quantitative PCR (qPCR), enzyme-linked immunosorbent assay (ELISA), and Western blot assay. miR-126 KD cells showed lower expression of interleukin 8 (IL-8) and CXCL1, both on the mRNA and protein levels, than did controls upon stimulation by heat-killed wild-type Porphyromonas gingivalis, live P. gingivalis protease-deficient mutant KDP128, and live Aggregatibacter actinomycetemcomitans In contrast, infection of miR-155 KD and miR-210 KD cells with the same organisms resulted in higher IL-8 and CXCL1 mRNA and protein expression. These effects appeared to be regulated by NF-κB, as suggested by altered transcription and/or phosphorylation status of components of the NF-κB system. Reduced neutrophil-like HL-60 cell chemotactic activity was observed in response to infection of miR-126 KD cells, indicating that miR-126 plays an important role in immune responses. Our findings indicate that specific miRNAs regulate the expression of inflammatory cytokines in human gingival epithelial cells in response to microbial infection.

Original languageEnglish
Pages (from-to)3282-3289
Number of pages8
JournalInfection and Immunity
Volume84
Issue number12
Early online date18 Nov 2016
DOIs
Publication statusPublished - Dec 2016

Keywords

  • Aggregatibacter actinomycetemcomitans
  • Cell Line
  • Chemotaxis
  • Cytokines/genetics
  • Gene Expression Regulation/immunology
  • Gene Knockdown Techniques
  • Gingiva/cytology
  • HL-60 Cells
  • Humans
  • Keratinocytes/metabolism
  • MicroRNAs/genetics
  • Porphyromonas gingivalis

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