Tumor Biology

, Volume 36, Issue 12, pp 9171–9177 | Cite as

Cancer cell-derived IL-8 induces monocytic THP1 cells to secrete IL-8 via the mitogen-activated protein kinase pathway

  • Yukina Nishio
  • Takahiro Gojoubori
  • Yasuhide Kaneko
  • Noriyoshi Shimizu
  • Masatake Asano
Research Article


Aberrant activity of transcription factors in oral squamous cell carcinoma (OSCC) results in the spontaneous secretion of various cytokines and chemokines. Among them, IL-8, owing to its angiogenic activity, promotes the growth of OSCCs. In the present study, we examined the role of IL-8 secreted by OSCCs, on the angiogenic activity of monocytic THP1 cells. Culture supernatant (Ca-sup) augmented IL-8 secretion by THP1 cells, which was found to be significantly reduced following the removal Ca9-22-derived IL-8 from the Ca-sup. IL-8 induction was regulated at the transcriptional level, because real-time PCR demonstrated the augmented IL-8 messenger RNA (mRNA) expression. We further performed the luciferase assay using the 5′-untranslated region of IL-8 gene. Contradictory to our speculations, luciferase activity was not augmented by Ca-sup stimulation. NF-κB-independent IL-8 induction was further confirmed by pre-treating THP1 cells with NF-κB-specific inhibitors. To elucidate the signaling pathway, THP1 was pre-treated with MEK inhibitors. The results demonstrated that pre-treatment of cells with MEK inhibitor drastically reduced IL-8 levels, suggesting the role of MEK. Moreover, Ca-sup was found to increase ERK1/2 phosphorylation in a time-dependent manner. These results indicated that OSCC-derived IL-8 appears to activate angiogenic activity in monocytes within the tumor microenvironment via the mitogen-activated protein kinase (MAPK) pathway.


Oral squamous cell carcinoma Interleukin-8 Macrophage Mitogen-activated protein kinase 



This work was supported by a grant of Strategic Research Base Development Program for Private Universities from the Ministry of Education, Culture, Sports, Science, and Technology, Japan (MEXT), 2010–2014 (S1001024); MEXT-supported program for the Strategic Research Foundation at Private Universities, 2013–2018; Sato Fund, Uemura Fund, and Grant from the Dental Research Center Nihon University School of Dentistry; and Nihon University Multidisciplinary Research Grant for 2015.


  1. 1.
    Molinolo AA, Amornphimoltham P, Squarize CH, Castilho RM, Patel V, Gutkind JS. Dysregulated molecular networks in head and neck carcinogenesis. Oral Oncol. 2009;45:324–34.CrossRefPubMedGoogle Scholar
  2. 2.
    Ondrey FG, Dong G, Sunwoo J, Chen Z, Wolf JS, Crowl-Bancroft CV, et al. Constitutive activation of transcription factors NF-(kappa)B, AP-1, and NF-IL6 in human head and neck squamous cell carcinoma cell lines that express pro-inflammatory and pro-angiogenic cytokines. Mol Carcinog. 1999;26:119–29.CrossRefPubMedGoogle Scholar
  3. 3.
    Bindhu OS, Ramadas K, Sebastian P, Pillai MR. High expression levels of nuclear factor kappa B and gelatinases in the tumorigenesis of oral squamous cell carcinoma. Head Neck. 2006;28:916–25.CrossRefPubMedGoogle Scholar
  4. 4.
    Sawhney M, Rohatgi N, Kaur J, Shishodia S, Sethi G, Gupta SD, et al. Expression of NF-kappaB parallels COX-2 expression in oral precancer and cancer: association with smokeless tobacco. Int J Cancer. 2007;120:2545–56.CrossRefPubMedGoogle Scholar
  5. 5.
    Mishra A, Bharti AC, Varghese P, Saluja D, Das BC. Differential expression and activation of NF-kappaB family proteins during oral carcinogenesis: role of high risk human papillomavirus infection. Int J Cancer. 2006;119:2840–50.CrossRefPubMedGoogle Scholar
  6. 6.
    Squarize CH, Castilho RM, Sriuranpong V, Pinto Jr DS, Gutkind JS. Molecular cross-talk between the NFkappaB and STAT3 signaling pathways in head and neck squamous cell carcinoma. Neoplasia. 2006;8:733–46.CrossRefPubMedPubMedCentralGoogle Scholar
  7. 7.
    Balkwill FR. The chemokine system and cancer. J Pathol. 2012;226:148–57.CrossRefPubMedGoogle Scholar
  8. 8.
    Richmond A. NF-κb, chemokine gene transcription and tumour growth. Nat Rev Immunol. 2002;2:664–74.CrossRefPubMedPubMedCentralGoogle Scholar
  9. 9.
    Shionome T, Endo S, Omagari D, Asano M, Toyoma H, Ishigami T, et al. Nickel ion inhibits nuclear factor-kappa B activity in human oral squamous cell carcinoma. PLoS ONE. 2013;8, e68257.CrossRefPubMedPubMedCentralGoogle Scholar
  10. 10.
    Watanabe H, Iwase M, Ohashi M, Nagumo M. Role of interleukin-8 secreted from human oral squamous cell carcinoma cell lines. Oral Oncol. 2002;38:670–9.CrossRefPubMedGoogle Scholar
  11. 11.
    Kessenbrock K, Plaks V, Werb Z. Matrix metalloproteinases: regulators of the tumor microenvironment. Cell. 2010;141:52–67.CrossRefPubMedPubMedCentralGoogle Scholar
  12. 12.
    Qian BZ, Pollard JW. Macrophage diversity enhances tumor progression and metastasis. Cell. 2010;141:39–51.CrossRefPubMedPubMedCentralGoogle Scholar
  13. 13.
    Condeelis J, Pollard JW. Macrophages: obligate partners for tumor cell migration, invasion, and metastasis. Cell. 2006;124:263–6.CrossRefPubMedGoogle Scholar
  14. 14.
    Lewis CE, Pollard JW. Distinct role of macrophages in different tumor microenvironments. Cancer Res. 2006;66:605–12.CrossRefPubMedGoogle Scholar
  15. 15.
    Li C, Shintani S, Terakado N, Nakashiro K, Hamakawa H. Infiltration of tumor-associated macrophages in human oral squamous cell carcinoma. Oncol Rep. 2002;9:1219–23.PubMedGoogle Scholar
  16. 16.
    Waugh DJ, Wilson C. The interleukin-8 pathway in cancer. Clin Cancer Res. 2008;14:6735–41.CrossRefPubMedGoogle Scholar
  17. 17.
    Zimmermann HW, Seidler S, Gassler N, Nattermann J, Luedde T, Trautwein C, et al. Interleukin-8 is activated in patients with chronic liver diseases and associated with hepatic macrophage accumulation in human liver fibrosis. PLoS One. 2011;6, e21381.CrossRefPubMedPubMedCentralGoogle Scholar
  18. 18.
    Omagari D, Mikami Y, Suguro H, Sunagawa K, Asano M, Sanuki E, et al. Poly I:C-induced expression of intercellular adhesion molecule-1 in intestinal epithelial cells. Clin Exp Immunol. 2009;156:294–302.CrossRefPubMedPubMedCentralGoogle Scholar
  19. 19.
    Hoffmann E, Dittrich-Breiholz O, Holtmann H, Kracht M. Multiple control of interleukin-8 gene expression. J Leukoc Biol. 2002;72:847–55.PubMedGoogle Scholar
  20. 20.
    Kerbel R, Folkman J. Clinical translation of angiogenesis inhibitors. Nat Rev Cancer. 2002;2:727–39.CrossRefPubMedGoogle Scholar
  21. 21.
    Torisu H, Ono M, Kiryu H, Furue M, Ohmoto Y, Nakayama J, et al. Macrophage infiltration correlates with tumor stage and angiogenesis in human malignant melanoma: possible involvement of TNFalpha and IL-1alpha. Int J Cancer. 2000;85:182–8.CrossRefPubMedGoogle Scholar
  22. 22.
    Schoppmann SF, Birner P, Stockl J, Kalt R, Ullrich R, Caucig C, et al. Tumor-associated macrophages express lymphatic endothelial growth factors and are related to peritumoral lymphangiogenesis. Am J Pathol. 2002;161:947–56.CrossRefPubMedPubMedCentralGoogle Scholar
  23. 23.
    Oppenheim JJ, Zachariae CO, Mukaida N, Matsushima K. Properties of the novel proinflammatory supergene “intercrine” cytokine family. Annu Rev Immunol. 1991;9:617–48.CrossRefPubMedGoogle Scholar
  24. 24.
    Campbell LM, Maxwell PJ, Waugh DJ. Rationale and means to target pro-inflammatory interleukin-8 (CXCL8) signaling in cancer. Pharmaceuticals (Basel). 2013;6:929–59.CrossRefGoogle Scholar
  25. 25.
    Hou Y, Ryu CH, Jun JA, Kim SM, Jeong CH, Jeun SS. IL-8 enhances the angiogenic potential of human bone marrow mesenchymal stem cells by increasing vascular endothelial growth factor. Cell Biol Int. 2014;38:1050–9.Google Scholar
  26. 26.
    Singh RK, Gutman M, Radinsky R, Bucana CD, Fidler IJ. Expression of interleukin 8 correlates with the metastatic potential of human melanoma cells in nude mice. Cancer Res. 1994;54:3242–7.PubMedGoogle Scholar
  27. 27.
    Kitadai Y, Haruma K, Sumii K, Yamamoto S, Ue T, Yokozaki H, et al. Expression of interleukin-8 correlates with vascularity in human gastric carcinomas. Am J Pathol. 1998;152:93–100.PubMedPubMedCentralGoogle Scholar
  28. 28.
    Fujimoto J, Sakaguchi H, Aoki I, Tamaya T. Clinical implications of expression of interleukin 8 related to angiogenesis in uterine cervical cancers. Cancer Res. 2000;60:2632–5.PubMedGoogle Scholar
  29. 29.
    Yatsunami J, Tsuruta N, Ogata K, Wakamatsu K, Takayama K, Kawasaki M, et al. Interleukin-8 participates in angiogenesis in non-small cell, but not small cell carcinoma of the lung. Cancer Lett. 1997;120:101–8.CrossRefPubMedGoogle Scholar
  30. 30.
    Chen Z, Malhotra PS, Thomas GR, Ondrey FG, Duffey DC, Smith CW, et al. Expression of proinflammatory and proangiogenic cytokines in patients with head and neck cancer. Clin Cancer Res. 1999;5:1369–79.PubMedGoogle Scholar
  31. 31.
    Balkwill F, Mantovani A. Inflammation and cancer: back to Virchow? Lancet. 2001;357:539–45.CrossRefPubMedGoogle Scholar

Copyright information

© International Society of Oncology and BioMarkers (ISOBM) 2015

Authors and Affiliations

  • Yukina Nishio
    • 1
  • Takahiro Gojoubori
    • 2
  • Yasuhide Kaneko
    • 2
  • Noriyoshi Shimizu
    • 3
  • Masatake Asano
    • 2
  1. 1.Division of Oral Structural and Functional BiologyNihon University Graduate School of DentistryTokyoJapan
  2. 2.Department of PathologyNihon University School of DentistryTokyoJapan
  3. 3.Department of OrthodonticsNihon University School of DentistryTokyoJapan

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