Poly I:C inhibits cell proliferation and enhances the growth inhibitory effect of paclitaxel in oral sqaumous cell carcinoma.

OBJECTIVE: Toll-like receptors (TLR) signaling has dual effect of promoting tumor progression and anti-cancer property. This study was designed to determine the effect of polyinosinic-polycytidilic acid (poly I:C), a TLR3 agonist, on the proliferation of oral cancer cells. MATERIALS AND METHODS: Human oral squamous cell carcinoma cell lines, YD-10B and YD-8, were used. TLRs expression was examined by RT-PCR and IL-8 production by poly I:C was examined by ELISA. Cell proliferation was determined by MTT assay. Flow cytometry and Western blot analysis were performed to determine the molecular mechanism of poly I:C-induced cell death. RESULTS: TLR3 was functionally expressed in YD-10B and YD-8 cells. Treatment of poly I:C inhibited the cell growth in a dose-dependent manner. Flow cytometry and Western blot analysis revealed that poly I:C induced apoptosis via a mitochondria-dependent pathway. In addition, combination treatment with poly I:C and paclitaxel more significantly inhibited cell proliferation compared with poly I:C or paclitaxel alone. CONCLUSIONS: Poly I:C effectively inhibits oral cancer cell proliferation and can be considered as a candidate to improve the inhibitory effect of anti-cancer drugs.

Full-field optical coherence microscopy for identifying live cancer cells by quantitative measurement of refractive index distribution.

The feasibility of identifying cancer cells by measuring the refractive index (RI) distribution across a single live cell with ultrahigh resolution full-field optical coherence microscopy (FF-OCM) is presented. The FF-OCM is utilized to quantify integral RI distributions of unmodified cells without any cell treatments and used as a biophysical indicator for diagnosing cell malignancy. Firstly, the physical thickness distribution of the cell adherent to a culture dish is measured by taking a series of 0.6 µm resolved en-face tomograms. Subsequently, from the en-face image of the bottom surface of the cell or the top surface of the dish, the phase gain image of the cell is extracted. Then, from these two measurements the axially averaged RI map of the cell is extracted. The implemented FF-OCM system had a 0.8 µm axial resolution and the phase measurement sensitivity of the system was around 124 mrad. With the system, RI maps of several living cell lines of normal and cancer cells were constructed and quantitatively analyzed. The experiments showed that cancer cells had higher RI than normal ones. This approach using the FF-OCM has significant potential for cancer diagnosis and dynamic cell analysis as in situ label-free biophysical assay.

Activation of TLR2 and TLR5 did not affect tumor progression of an oral squamous cell carcinoma, YD-10B cells.

BACKGROUND: Toll-like receptors (TLRs) signaling has been found to promote cell proliferation, invasiveness, and angiogenesis in a variety of cancers. This study was performed to examine whether TLR signaling is involved in tumor progression of an oral squamous cell carcinoma, YD-10B cells. METHODS: TLRs expression was examined by reverse transcription-polymerase chain reaction (RT-PCR) in YD-10B cells. Interleukin (IL)-6 and IL-8 production by YD-10B cells in response to various TLR agonists was examined by ELISA. Cell viability and proliferation was determined by colorimetric MTT and Bromodeoxyuridine (BrdU) assay. The effect of TLR agonists on invasiveness was determined by migration and invasion assay using commercial kits. mRNA expression of vascular endothelial growth factor (VEGF) was also evaluated by RT-PCR. RESULTS: All tested TLRs including TLR2, 3, 4, 5, 7, and 9 were expressed in YD-10B cells. IL-6 and IL-8 production was increased by Pam(3) CSK(4) , flagellin, Poly I:C, and imiquimod, but not lipopolysaccharide (LPS). Porphyromonas gingivalis LPS (Pg LPS) also led to increase of IL-8 production. However, Pam(3) CSK(4,) flagellin, and Pg LPS did not affect cell proliferation, migration, invasion, and gene expression of VEGF in YD-10B cells. CONCLUSION: These findings indicated that TLR activation by bacterial molecules may not affect tumor progression of YD-10B cells.

NOD1 and NOD2 stimulation triggers innate immune responses of human periodontal ligament cells.

Nod-like receptors (NLRs) are cytosolic sensors for microbial molecules. Νucleotide-binding oligomerization domain (NOD)1 and NOD2 recognize the peptidoglycan derivatives, meso-diaminopimelic acid (meso-DAP) and muramyl dipeptide (MDP), respectively, and trigger host innate immune responses. In the present study, we examined the function of NOD1 and NOD2 on innate immune responses in human periodontal ligament (PDL) cells. The gene expression of NOD1 and NOD2 was examined by RT-PCR. IL-6 and IL-8 production in culture supernatants was measured by ELISA. Western blot analysis was performed to determine the activation of NF-κB and MAPK in response to Tri-DAP and MDP. The genes of NOD1 and NOD2 appeared to be expressed in PDL cells. Although the levels of NOD2 expression were weak in intact cells, MDP stimulation increased the gene expression of NOD2 in PDL cells. Tri-DAP and MDP led to the production of IL-6 and IL-8 and the activation of NF-κB and MAPK in PDL cells. Toll-like receptor (TLR) stimulation led to increased gene expression of NOD1 and NOD2 in PDL cells. Pam3CSK4 (a TLR2 agonist) and IFN-γ synergized with Tri-DAP and MDP to produce IL-8 and IL-6 in PDL cells. Our results indicate that NOD1 and NOD2 are functionally expressed in human PDL cells and can trigger innate immune responses.