The secretion of IL-6 by CpG-ODN-treated cancer cells promotes T-cell immune responses partly through the TLR-9/AP-1 pathway in oral squamous cell carcinoma

Ruan,Min; Thorn,Katherine; Liu,Shengwen; Li,Siyi; Yang,Wenjun; Zhang,Chunye; Zhang,Chenping

doi:10.3892/ijo.2014.2356

The secretion of IL-6 by CpG-ODN-treated cancer cells promotes T-cell immune responses partly through the TLR-9/AP-1 pathway in oral squamous cell carcinoma

Authors:
- Min Ruan
- Katherine Thorn
- Shengwen Liu
- Siyi Li
- Wenjun Yang
- Chunye Zhang
- Chenping Zhang
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Affiliations: Department of Oral and Maxillofacial-Head and Neck Oncology, Ninth People's Hospital, Shanghai Jiao Tong University School of Medicine, Shanghai Key Laboratory of Stomatology, Shanghai 200011, P.R. China , Department of Surgery, Memorial Sloan-Kettering Cancer Center, New York, NY 10065, USA, Department of Oral Pathology, Ninth People's Hospital, Shanghai Jiao Tong University School of Medicine, Shanghai Key Laboratory of Stomatology, Shanghai 200011, P.R. China
Published online on: March 21, 2014 https://doi.org/10.3892/ijo.2014.2356
Pages: 2103-2110

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Abstract

Increasing evidence suggests that communication between tumor and immune cells can alter the tumor microenvironment in ways that promote tumor development. The purpose of this study was to characterize the immune response elicited by TLR-9-activated OSCC cells, to identify the cytokines involved in the signaling pathway and to elucidate the molecular mechanism of this pathway in OSCC cells. MTS, flow cytometry and ELISA assay were used to evaluate T-cell immune responses, cancer cell proliferation and pro-inflammatory cytokine secretion, respectively. Western blot analysis, EMSA and ChIP assay were employed to detect the activity of the NF-κB and AP-1 signaling pathways. A marked response was observed when T-cells were co-cultured with supernatants from CpG-ODN-treated OSCC cells. This response was characterized by increased CD4+ and CD8+ T-cell proliferation and an increase in IFN-γ production by the CD4+ T-cell population. Treatment of OSCC cells with CpG-ODN resulted in an increase in IL-6 secretion as well as an increase in AP-1 binding activity to the IL-6 promoter. Moreover, blockage of the TLR-9/AP-1 pathway significantly decreased IL-6 expression and T-cell immune response. In human OSCC, the TLR-9 pathway, when stimulated by CpG-ODNs, promotes a T-cell immune response mediated by AP-1-activated IL-6 secretion. Although the complete molecular mechanism has yet to be understood, these findings provide evidence linking tumor cell activities to immune system responses. In addition, the TLR-9/AP-1/IL-6 pathway provides new therapeutic targets for the prevention and treatment of OSCC.

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1.	Parkin DM, Bray F, Ferlay J and Pisani P: Global cancer statistics, 2002. CA Cancer J Clin. 55:74–108. 2005. View Article : Google Scholar
2.	Ferlay J, Shin HR, Bray F, Forman D and Mathers C: GLOBOCAN 2008, cancer incidence and mortality worldwide: IARC Cancer Base No. 10. International Agency for Research on Cancer. http://globocan.iarc.fr. Accessed July 20, 2012.
3.	Forastiere AA, Goepfert H, Maor M, Pajak TF, Weber R, et al: Concurrent chemotherapy and radiotherapy for organ preservation in advanced laryngeal cancer. N Engl J Med. 349:2091–2098. 2003. View Article : Google Scholar : PubMed/NCBI
4.	Melief CJ: Cancer: immune pact with the enemy. Nature. 450:803–804. 2007. View Article : Google Scholar : PubMed/NCBI
5.	Colotta F, Allavena P, Sica A, Garlanda C and Mantovani A: Cancer-related inflammation, the seventh hallmark of cancer: links to genetic instability. Carcinogenesis. 30:1073–1081. 2009. View Article : Google Scholar : PubMed/NCBI
6.	Coussens LM and Werb Z: Inflammation and cancer. Nature. 420:860–867. 2002. View Article : Google Scholar : PubMed/NCBI
7.	Greten FR, Eckmann L, Greten TF, Park JM, Li ZW, et al: IKKbeta links inflammation and tumorigenesis in a mouse model of colitis-associated cancer. Cell. 118:285–296. 2004. View Article : Google Scholar : PubMed/NCBI
8.	Maeda S, Chang L, Li ZW, Luo JL, Leffert H, et al: IKKbeta is required for prevention of apoptosis mediated by cell-bound but not by circulating TNFalpha. Immunity. 19:725–737. 2003. View Article : Google Scholar
9.	Rakoff-Nahoum S, Paglino J, Eslami-Varzaneh F, Edberg S and Medzhitov R: Recognition of commensal microflora by toll-like receptors is required for intestinal homeostasis. Cell. 118:229–241. 2004. View Article : Google Scholar
10.	El-Omar EM, Ng MT and Hold GL: Polymorphisms in Toll-like receptor genes and risk of cancer. Oncogene. 27:244–252. 2008. View Article : Google Scholar : PubMed/NCBI
11.	O'Neill LA: Signal transduction pathways activated by the IL-1 receptor/Toll-like receptor super family. Curr Top Microbiol Immunol. 270:47–61. 2002.
12.	Takeda K and Akira S: Toll-like receptors in innate immunity. Int Immunol. 17:1–14. 2005. View Article : Google Scholar
13.	Ruan M, Zun Z, Siyi L, Wenjun Y, Lizheng W, et al: Increased expression of Toll-like receptor-9 has close relation with tumour cell proliferation in oral squamous cell carcinoma. Arch Oral Biol. 56:877–884. 2011. View Article : Google Scholar : PubMed/NCBI
14.	Min R, Siyi L, Wenjun Y, Shengwen L, Ow A, et al: Toll-like receptor-9 agonists increase cyclin D1 expression partly through activation of activator protein-1 in human oral squamous cell carcinoma cells. Cancer Sci. 103:1938–1945. 2012. View Article : Google Scholar : PubMed/NCBI
15.	Sdek P, Zhang ZY, Cao J, Pan HY, Chen WT, et al: Alteration of cell-cycle regulatory proteins in human oral epithelial cells immortalized by HPV16 E6 and E7. Int J Oral Maxillofac Surg. 35:653–657. 2006. View Article : Google Scholar : PubMed/NCBI
16.	Zhong LP, Pan HY, Zhou XJ, Ye DX, Zhang L, et al: Characteristics of a cancerous cell line, HIOEC-B(a)P-96, induced by benzo(a)pyrene from human immortalized oral epithelial cell line. Arch Oral Biol. 53:443–452. 2008. View Article : Google Scholar : PubMed/NCBI
17.	Ruan M, Ji T, Yang WJ, Duan WH, Zhou XJ, et al: Growth inhibition and induction of apoptosis in human oral squamous cell carcinoma Tca-8113 cell lines by Shikonin was partly through the inactivation of NF-κB pathway. Phytother Res. 22:407–415. 2008.PubMed/NCBI
18.	Kaomongkolgit R, Cheepsunthorn P, Pavasant P and Sanchavanakit N: Iron increases MMP-9 expression through activation of AP-1 via ERK/Akt pathway in human head and neck squamous carcinoma cells. Oral Oncol. 44:587–594. 2008. View Article : Google Scholar : PubMed/NCBI
19.	Toualbi-Abed K, Daniel F, Güller MC, Legrand A, Mauriz JL, et al: Jun D cooperates with p65 to activate the proximal κB site of the cyclin D1 promoter: role of PI3K/PDK-1. Carcinogenesis. 29:536–543. 2008.PubMed/NCBI
20.	Woods KV, El-Naggar A, Clayman GL and Grimm EA: Variable expression of cytokines in human head and neck squamous cell carcinoma cell lines and consistent expression in surgical specimens. Cancer Res. 58:3132–3141. 1998.PubMed/NCBI
21.	St John MA, Li Y, Zhou X, Denny P, Ho CM, et al: Interleukin 6 and interleukin 8 as potential biomarkers for oral cavity and oropharyngeal squamous cell carcinoma. Arch Otolaryngol Head Neck Surg. 130:929–935. 2004.PubMed/NCBI
22.	Nagata M, Fujita H, Ida H, Hoshina H, Inoue T, et al: Identification of potential biomarkers of lymph node metastasis in oral squamous cell carcinoma by cDNA microarray analysis. Int J Cancer. 106:683–689. 2003. View Article : Google Scholar : PubMed/NCBI
23.	Okamoto M, Hiura K, Ohe G, Ohba Y, Terai K, et al: Mechanism for bone invasion of oral cancer cells mediated by interleukin-6 in vitro and in vivo. Cancer. 89:1966–1975. 2000. View Article : Google Scholar : PubMed/NCBI
24.	De Schutter H, Landuyt W, Verbeken E, Goethals L, Hermans R, et al: The prognostic value of the hypoxia markers CA IX and GLUT 1 and the cytokines VEGF and IL 6 in head and neck squamous cell carcinoma treated by radiotherapy +/− chemotherapy. BMC Cancer. 5:422005.PubMed/NCBI
25.	Okamoto M, Lee C and Oyasu R: Interleukin-6 as a paracrine and autocrine growth factor in human prostatic carcinoma cells in vitro. Cancer Res. 57:141–146. 1997.PubMed/NCBI
26.	Smith HA and Kang Y: The metastasis-promoting roles of tumor-associated immune cells. J Mol Med. 91:411–429. 2013. View Article : Google Scholar : PubMed/NCBI
27.	Beetz A, Peter RU, Oppel T, Kaffenberger W, Rupec RA, et al: NF-kappaB and AP-1 are responsible for inducibility of the IL-6 promoter by ionizing radiation in HeLa cells. Int J Radiat Biol. 76:1443–1453. 2000. View Article : Google Scholar : PubMed/NCBI
28.	Romano M, Sironi M, Toniatti C, Polentarutti N, Fruscella P, et al: Role of IL-6 and its soluble receptor in induction of chemokines and leukocyte recruitment. Immunity. 6:315–325. 1997. View Article : Google Scholar : PubMed/NCBI
29.	Mishra A, Bharti AC, Saluja D and Das BC: Transactivation and expression patterns of Jun and Fos/AP-1 super-family proteins in human oral cancer. Int J Cancer. 126:819–829. 2010.PubMed/NCBI
30.	Rao SK, Pavicevic Z, Du Z, Kim JG, Fan M, Jiao Y, et al: Pro-inflammatory genes as biomarkers and therapeutic targets in oral squamous cell carcinoma. J Biol Chem. 285:32512–32521. 2010. View Article : Google Scholar : PubMed/NCBI
31.	Lin CM, Chen YH, Ma HP, Wang BW, Chiu JH, et al: Silibinin inhibits the invasion of IL-6-stimulated colon cancer cells via selective JNK/AP-1/MMP-2 modulation in vitro. J Agric Food Chem. 60:12451–12457. 2012. View Article : Google Scholar : PubMed/NCBI
32.	Houghton AN, Uchi H and Wolchok JD: The role of the immune system in early epithelial carcinogenesis: B-ware the double-edged sword. Cancer Cell. 7:403–405. 2005. View Article : Google Scholar : PubMed/NCBI
33.	Huang B, Zhao J, Unkeless JC, Feng ZH and Xiong H: TLR signaling by tumor and immune cells: a double-edged sword. Oncogene. 27:218–224. 2008. View Article : Google Scholar
34.	Anders CK, Hsu DS, Broadwater G, Acharya CR, Foekens JA, et al: Young age at diagnosis correlates with worse prognosis and defines a subset of breast cancers with shared patterns of gene expression. J Clin Oncol. 26:3324–3330. 2008. View Article : Google Scholar : PubMed/NCBI
35.	Hirsh V, Paz-Ares L, Boyer M, Rosell R, Middleton G, et al: Randomized phase III trial of paclitaxel/carboplatin with or without PF-3512676 (Toll-like receptor 9 agonist) as first-line treatment for advanced non-small-cell lung cancer. J Clin Oncol. 29:2667–2674. 2011. View Article : Google Scholar : PubMed/NCBI