TLR5 involvement in attenuated IL-8 production in nuclear decorin silenced oral mucosal dysplastic keratinocytes and squamous cell carcinoma

Nyla  Dil; Abhijit G.  Banerjee

doi:10.4236/ajmb.2013.34032

AJMB Vol.3 No.4 , October 2013

TLR5 involvement in attenuated IL-8 production in nuclear decorin silenced oral mucosal dysplastic keratinocytes and squamous cell carcinoma

Nyla Dil^*, Abhijit G. Banerjee

Abstract: Head and neck cancer is one of the most prevalent cancers in the world. Roughly half of these malignancies originate from oral mucosa and constitute Oral squamous cell carcinomas. Despite many advances in diagnostic and therapeutic regimens, five year survival rate remains at roughly 50 %, indicating the need for in depth understanding of the oral squamous cell carcinoma immunobiology. We have previously shown that in human dysplastic oral keratinocytes (DOK) and malignant squamous cells carcinoma (SCC-25), multifunctional proteoglycan decorin is aberrantly expressed and localized in the nucleus bound to nuclear EGFR. In vitro nuclear decorin knockdown significantly reduced IL-8 and IL8-dependent migration, invasion and angiogenesis in these cells. Since toll-like receptor (TLR) signalling leads to IL-8 production we examined here if these receptors play a role in decorin silencing mediated reduction in IL-8 levels. We have used immunological and molecular techniques to study toll-like receptors involvement in attenuated IL-8 production in nuclear decorin silenced (stable knockdown) oral mucosal dysplastic keratinocytes and squamous carcinoma cells. We show that nuclear decorin silenced DOK and SCC-25 cells show marked diminution of TLR5 mRNA and protein expression compared with respective controls that translated into loss of function in response to appropriate TLR ligand. In these mucosal oral epithelia, decorin stable knockdown significantly down-regulated IL-8 production following activation with TLR5 ligand flagellin. These data suggest that decorin silencing interferes with IL-8 production, in part, by altering TLR5 expression and signaling in dysplastic and malignant oral epithelia. This study highlights the significance of TLR5 expression and signaling in mucosal cancers.

Keywords: Oral Cancer, Proteoglycans, Flagellin, DAMP, Mucosal Cancer, TLR5, PAMP, IL-8, SLRP, Nuclear decorin

Cite this paper: Dil, N. and Banerjee, A.G. (2013) TLR5 involvement in attenuated IL-8 production in nuclear decorin silenced oral mucosal dysplastic keratinocytes and squamous cell carcinoma. American Journal of Molecular Biology, 3, 248-254. doi: 10.4236/ajmb.2013.34032.

References

[1] Jemal, A., Siegel, R., Ward, E., Hao, Y.P., Xu J.Q., Murray, T. and Thun, M.J. (2008) Cancer statistics, 2008. CA: A Cancer Journal for Clinicians, 58, 71-96.
http://dx.doi.org/10.3322/CA.2007.0010

[2] Jemal, A., Center, M.M., Ward, E. and Thun, M.J. (2009) Cancer occurrence. Methods in Molecular Biology, 471, 3-29. http://dx.doi.org/10.1007/978-1-59745-416-2_1

[3] Parkin, D.M., Bray, F., Ferlay, J. and Pisani, P. (2005) Global cancer statistics, 2002. CA: A Cancer Journal for Clinicians, 55, 74-108.
http://dx.doi.org/10.3322/canjclin.55.2.74

[4] Dobrossy, L. (2005) Epidemiology of head and neck cancer: Magnitude of the problem. Cancer and Metastasis Reviews, 24, 9-17.
http://dx.doi.org/10.1007/s10555-005-5044-4

[5] Ramqvist, T. and Dalianis, T. (2010) Oropharyngeal cancer epidemic and human papillomavirus. Emerging Infectious Diseases, 16, 1671-1677.
http://dx.doi.org/10.3201/eid1611.100452

[6] Medzhitov, R., Preston-Hurlburt, P. and Janeway, Jr. C.A. (1997) A human homologue of the Drosophila Toll protein signals activation of adaptive immunity. Nature, 388, 394-397.
http://dx.doi.org/10.1038/41131

[7] Takeda, K., Kaisho, T. and Akira, S. (2003) Toll-like receptors. Annual Review of Immunology, 21, 335-376.
http://dx.doi.org/10.1146/annurev.immunol.21.120601.141126

[8] Akira, S., Uematsu, S. and Takeuchi, O. (2006) Pathogen recognition and innate immunity. Cell, 124, 783-801.
http://dx.doi.org/10.1016/j.cell.2006.02.015

[9] Furrie, E., et al. (2005) Toll-like receptors-2, -3 and -4 expression patterns on human colon and their regulation by mucosal-associated bacteria. Immunology, 115, 565-574.
http://dx.doi.org/10.1111/j.1365-2567.2005.02200.x

[10] Kelly, M.G., et al. (2006) TLR-4 signaling promotes tumor growth and paclitaxel chemoresistance in ovarian cancer. Cancer Research, 66, 3859-3868.
http://dx.doi.org/10.1158/0008-5472.CAN-05-3948

[11] Lee, J.W., et al. (2007) Increased toll-like receptor 9 expression in cervical neoplasia. Molecular Carcinogenesis, 46, 941-947. http://dx.doi.org/10.1002/mc.20325

[12] Park, J.H., et al. (2011) Toll-like receptor 5 activation promotes migration and invasion of salivary gland adenocarcinoma. Journal of Oral Pathology & Medicine, 40, 187-193.
http://dx.doi.org/10.1111/j.1600-0714.2010.00929.x

[13] He, W., et al. (2007) TLR4 signaling promotes immune escape of human lung cancer cells by inducing immunosuppressive cytokines and apoptosis resistance. Molecular Immunology, 44, 2850-2859.
http://dx.doi.org/10.1016/j.molimm.2007.01.022

[14] Droemann, D., et al. (2005) Human lung cancer cells express functionally active Toll-like receptor 9. Respiratory Research, 6, 1. http://dx.doi.org/10.1186/1465-9921-6-1

[15] Koch, A.E., et al. (1992) Interleukin-8 as a macrophage-derived mediator of angiogenesis. Science, 258, 1798-801. http://dx.doi.org/10.1126/science.1281554

[16] Strieter, R.M., et al. (1995) Role of C-X-C chemokines as regulators of angiogenesis in lung cancer. Journal of Leukocyte Biology, 57, 752-762.

[17] Murdoch, C., et al. (1999) Cxc chemokine receptor expression on human endothelial cells. Cytokine, 11, 704-712. http://dx.doi.org/10.1006/cyto.1998.0465

[18] Dil, N. and Banerjee, A.G. (2011) A role for aberrantly expressed nuclear localized decorin in migration and invasion of dysplastic and malignant oral epithelial cells. Head & Neck Oncology, 3, 44.
http://dx.doi.org/10.1186/1758-3284-3-44

[19] Lalla, R.V., et al. (2001) Fibrin induces IL-8 expression from human oral squamous cell carcinoma cells. Oral Oncology, 37, 234-242.
http://dx.doi.org/10.1016/S1368-8375(00)00090-7

[20] Dil, N.and Banerjee, A.G. (2012) Knockdown of aberrantly expressed nuclear localized decorin attenuates tumour angiogenesis related mediators in oral cancer progression model in vitro. Head & Neck Oncology, 4, 11.
http://dx.doi.org/10.1186/1758-3284-4-11

[21] Banerjee, A.G., et al. (2003) Aberrant expression and localization of decorin in human oral dysplasia and squamous cell carcinoma. Cancer Research, 63, 7769-7776.

[22] Hu, L., et al. (1991) Abnormal expression of retinoic acid receptors and keratin 19 by human oral and epidermal squamous cell carcinoma cell lines. Cancer Research, 51, 3972-3981.

[23] Hsu, S. et al. (2002) Transforming growth factor beta 1 dysregulation in a human oral carcinoma tumour progression model. Cell Proliferation, 35, 183-192.
http://dx.doi.org/10.1046/j.1365-2184.2002.00237.x

[24] Yu, L. and Chen, S. (2008) Toll-like receptors expressed in tumor cells: Targets for therapy. Cancer Immunology, Immunotherapy, 57, 1271-1278.
http://dx.doi.org/10.1007/s00262-008-0459-8

[25] Neill, T., et al. (2012) Decorin: A guardian from the matrix. American Journal of Pathology, 181, 380-387.
http://dx.doi.org/10.1016/j.ajpath.2012.04.029

[26] Zafiropoulos, A., et al. (2008) Decorin-induced growth inhibition is overcome through protracted expression and activation of epidermal growth factor receptors in osteosarcoma cells. Molecular Cancer Research, 6, 785-794. http://dx.doi.org/10.1158/1541-7786.MCR-07-0165

[27] Zafiropoulos, A. and Tzanakakis, G.N. (2008) Decorin-mediated effects in cancer cell biology. Connective Tissue Research, 49, 244-248.
http://dx.doi.org/10.1080/03008200802147746

[28] Santra, M., et al. (2008) Ectopic decorin expression up-regulates VEGF expression in mouse cerebral endothelial cells via activation of the transcription factors Sp1, HIF1alpha, and Stat 3. Journal of Neurochemistry, 105, 324-337.
http://dx.doi.org/10.1111/j.1471-4159.2007.05134.x

[29] Schmausser, B., et al. (2005) Toll-like receptors TLR4, TLR5 and TLR9 on gastric carcinoma cells: An implication for interaction with Helicobacter pylori. International Journal of Medical Microbiology, 295, 179-185.
http://dx.doi.org/10.1016/j.ijmm.2005.02.009

[30] Pimentel-Nunes, P., et al. (2011) Increased expression of Toll-like receptors (TLR) 2, 4 and 5 in gastric dysplasia. Pathology & Oncology Research, 17, 677-683.
http://dx.doi.org/10.1007/s12253-011-9368-9

[31] Kim, W.Y., et al. (2008) Increased expression of Tolllike receptor 5 during progression of cervical neoplasia. International Journal of Gynecological Cancer, 18, 300-305.
http://dx.doi.org/10.1111/j.1525-1438.2007.01008.x

[32] Song, E.J., et al. (2011) Flagellin promotes the proliferation of gastric cancer cells via the Toll-like receptor 5. International Journal of Medical Microbiology, 28, 115-119.

[33] Kauppila, J.H., et al. (2013) Toll-like receptor 5 (TLR5) expression is a novel predictive marker for recurrence and survival in squamous cell carcinoma of the tongue. British Journal of Cancer, 108, 638-643.
http://dx.doi.org/10.1038/bjc.2012.589

[34] Schaefer, L., et al. (2005) The matrix component biglycan is proinflammatory and signals through Toll-like receptors 4 and 2 in macrophages. Journal of Clinical Investigation, 115, 2223-2233.
http://dx.doi.org/10.1172/JCI23755

[35] Frey, H., et al. (2013) Biological interplay between proteoglycans and their innate immune receptors in inflammation. FEBS Journal, 280, 2165-2179.

[36] Chen, G.Y. and Nunez, G. (2010) Sterile inflammation: Sensing and reacting to damage. Nature Reviews Immunology, 10, 826-837. http://dx.doi.org/10.1038/nri2873