Increasing evidence suggests that malignant transformation can result from
chronic infection, and
Toll-like receptors (TLRs) may play an important role in this process. We have previously reported that the increased expression of TLR-9 is associated with
tumor cell proliferation in
oral cancer. However, the mechanisms involved have not been elucidated. The aim of this study was to investigate whether CpG-
oligodeoxynucleotides (
CpG-ODN), a special TLR-9 agonist, is able to exert the proliferation-promoting effect in human
oral squamous cell carcinoma (OSCC), and to explore the possible underlying molecular mechanism. Flow cytometry, MTT, and colony formation assay were used to evaluate cell proliferation and cell cycle distribution. The
mRNA and
protein levels were analyzed by quantitative RT-PCR and Western blot assay.
Luciferase reporter gene, EMSA, and ChIP assays were used to detect the activity of
activator protein-1 (AP-1) and nuclear factor-κB (NF-κB) in HB cells. Results showed that
CpG-ODN could stimulate proliferation of HB cells in a dose- and time-dependent manner with a promoted G(1) /S cell cycle progression. Increased
cyclin D1 expression was detected in the nuclear region after
CpG-ODN treatment. Moreover,
CpG-ODN promoted nuclear translocation and activation of
AP-1, which appeared to be required for TLR-9-mediated
cyclin D1 expression and subsequently cell proliferation, but seemed to have little impact on NF-κB activity. Our results indicate that
CpG-ODN stimulates
tumor cell proliferation through TLR-9-mediated AP-1-activated
cyclin D1 expression in OSCC HB cells. Pharmacologic inhibition of the TLR-9/AP-1/
cyclin D1 pathway may be a new therapeutic approach for prevention as well as treatment of OSCC.