Clinical & Experimental Metastasis

, Volume 28, Issue 8, pp 887–897 | Cite as

Interleukin-6 receptor enhances early colonization of the murine omentum by upregulation of a mannose family receptor, LY75, in ovarian tumor cells

  • Premkumar Vummidi Giridhar
  • Holly M. Funk
  • Catherine A. Gallo
  • Aleksey Porollo
  • Carol A. Mercer
  • David R. Plas
  • Angela F. Drew
Research Paper


One of the earliest metastatic events in human ovarian cancer, tumor spread to the omentum, may be influenced by expression of interleukin 6 (IL6) and its cognate receptor (IL6Rα). Previous reports have shown that IL6 and IL6Rα expression is elevated in the serum and ascites of patients with ovarian cancer and that this can influence in vitro processes such as cell survival, proliferation and migration. In this study, overexpression of IL6Rα, and to a lesser extent IL6, enhanced tumor growth on the omentum. Moreover, adherence to plastic and to peritoneal extracellular matrix components was enhanced in tumor cells overexpressing IL6 or IL6Rα. Host production of IL6 and IL6Rα was also sufficient to influence tumor adherence to the omentum. Expression of LY75/CD205/DEC205, a collagen-binding mannose family receptor, was directly influenced by IL6Rα expression. Blocking LY75 with antibody reduced the adherence of tumor cells overexpressing IL6Rα to matrices in vitro and to the omentum. The association between IL6Rα expression and LY75 expression has not been previously reported, and the promotion of cellular adherence is a novel role for LY75. These studies indicate that overexpression of LY75 may be an additional mechanism by which IL6 signaling influences the progression of ovarian cancer, and suggests that blocking LY75 could be a valuable clinical strategy for reducing the early metastasis of ovarian cancer.


LY75 CD205 Interleukin-6 Ovarian cancer Omentum Metastasis 





IL6 receptor alpha


Janus-associated kinase


Signal transducer and activator of transcription


Extracellular signal-regulated kinase


Dulbecco’s modified Eagles medium


Red fluorescent protein

Supplementary material

10585_2011_9420_MOESM1_ESM.docx (14 kb)
Supplementary material 1 (DOCX 14 kb)
10585_2011_9420_MOESM2_ESM.tif (6.3 mb)
Supplemental Fig. 1Altered expression of IL6 and IL6Rα in Skov-3 cells. Q-PCR confirms altered mRNA expression in 2 variant Skov-3 cell lines (A and B) with overexpression (A) or knockdown shRNA (B) constructs for human IL6 (left panels) or IL6Rα (right panels). Data are expressed as fold change compared with cells expressing empty (control) vector (CV) or a scrambled sequence (pScramble). (C) Increased protein expression of IL6Rα was detected by ELISA of conditioned media collected from Skov-3 (1000 cells/well; 7 days). *p < 0.05. (D) Representative in vivo fluorescent images of omenta 48 h after injection of ES-2 cells. Arrows indicate the omentum (TIFF 6451 kb)
10585_2011_9420_MOESM3_ESM.tif (10.4 mb)
Supplemental Fig. 2IL6 and IL6Rα expression increase proliferative activity but not cell migration. (A) Expression of IL6 or IL6Rα was associated with increased proliferation/survival as assessed by MTT assay and (B) increased anchorage-independent growth on soft agar. (C) There were no differences in anoikis rates after 24 hours incubation on polyHEMA-coated plates among control cells and cells with altered expression of IL6 or IL6Rα. (D) The migration of cells through tissue culture inserts with 8 μm pores was not different for cells expressing control vector or IL6 or IL6Rα overexpression vectors. *p < 0.05 (TIFF 10627 kb)
10585_2011_9420_MOESM4_ESM.tif (2 mb)
Supplemental Fig. 3Addition of exogenous IL6 and IL6Rα increases adherence of tumor cells. Exogenous soluble IL6 (ng/mL) and IL6Rα (μg/mL) were added at the concentrations indicated to ES-2 cells expressing empty vector (CV) or IL6 or IL6Rα overexpression vectors, and adherence to plastic was assessed after 30 minutes (TIFF 2044 kb)


  1. 1.
    Rath KS et al (2010) Expression of soluble interleukin-6 receptor in malignant ovarian tissue. Am J Obstet Gynecol 203(3):230.e1–230.e8CrossRefGoogle Scholar
  2. 2.
    Jiang W et al (1995) The receptor DEC-205 expressed by dendritic cells and thymic epithelial cells is involved in antigen processing. Nature 375(6527):151–155PubMedCrossRefGoogle Scholar
  3. 3.
    Kato M et al (1998) cDNA cloning of human DEC-205, a putative antigen-uptake receptor on dendritic cells. Immunogenetics 47(6):442–450PubMedCrossRefGoogle Scholar
  4. 4.
    Kato M et al (2006) Expression of human DEC-205 (CD205) multilectin receptor on leukocytes. Int Immunol 18(6):857–869PubMedCrossRefGoogle Scholar
  5. 5.
    McKay PF et al (1998) The gp200-MR6 molecule which is functionally associated with the IL-4 receptor modulates B cell phenotype and is a novel member of the human macrophage mannose receptor family. Eur J Immunol 28(12):4071–4083PubMedCrossRefGoogle Scholar
  6. 6.
    Cheong C et al (2010) Improved cellular and humoral immune responses in vivo following targeting of HIV Gag to dendritic cells within human anti-human DEC205 monoclonal antibody. Blood 116(19):3828–3838PubMedCrossRefGoogle Scholar
  7. 7.
    Gurer C et al (2008) Targeting the nuclear antigen 1 of Epstein-Barr virus to the human endocytic receptor DEC-205 stimulates protective T-cell responses. Blood 112(4):1231–1239PubMedCrossRefGoogle Scholar
  8. 8.
    Shrimpton RE et al (2009) CD205 (DEC-205): a recognition receptor for apoptotic and necrotic self. Mol Immunol 46(6):1229–1239PubMedCrossRefGoogle Scholar
  9. 9.
    al-Tubuly AA et al (1996) Differential expression of gp200-MR6 molecule in benign hyperplasia and down-regulation in invasive carcinoma of the breast. Br J Cancer 74(7):1005–1011PubMedCrossRefGoogle Scholar
  10. 10.
    Tungekar MF, Gatter KC, Ritter MA (1996) Bladder carcinomas and normal urothelium universally express gp200-MR6, a molecule functionally associated with the interleukin 4 receptor (CD 124). Br J Cancer 73(4):429–432PubMedCrossRefGoogle Scholar
  11. 11.
    Robinson-Smith TM et al (2007) Macrophages mediate inflammation-enhanced metastasis of ovarian tumors in mice. Cancer Res 67(12):5708–5716PubMedCrossRefGoogle Scholar
  12. 12.
    Mosmann T (1983) Rapid colorimetric assay for cellular growth and survival: application to proliferation and cytotoxicity assays. J Immunol Methods 65(1–2):55–63PubMedCrossRefGoogle Scholar
  13. 13.
    Sood AK et al (2010) Adrenergic modulation of focal adhesion kinase protects human ovarian cancer cells from anoikis. J Clin Invest 120(5):1515–1523PubMedCrossRefGoogle Scholar
  14. 14.
    McFarland-Mancini MM et al (2010) Differences in wound healing in mice with deficiency of IL-6 versus IL-6 receptor. J Immunol 184(12):7219–7228PubMedCrossRefGoogle Scholar
  15. 15.
    Khan SM et al (2010) In vitro metastatic colonization of human ovarian cancer cells to the omentum. Clin Exp Metastasis 27(3):185–196PubMedCrossRefGoogle Scholar
  16. 16.
    Bild AH et al (2006) Oncogenic pathway signatures in human cancers as a guide to targeted therapies. Nature 439(7074):353–357PubMedCrossRefGoogle Scholar
  17. 17.
    Hendrix ND et al (2006) Fibroblast growth factor 9 has oncogenic activity and is a downstream target of Wnt signaling in ovarian endometrioid adenocarcinomas. Cancer Res 66(3):1354–1362PubMedCrossRefGoogle Scholar
  18. 18.
    Hagiwara A et al (1993) Milky spots as the implantation site for malignant cells in peritoneal dissemination in mice. Cancer Res 53(3):687–692PubMedGoogle Scholar
  19. 19.
    Krist LF et al (1998) Milky spots in the greater omentum are predominant sites of local tumour cell proliferation and accumulation in the peritoneal cavity. Cancer Immunol Immunother 47(4):205–212PubMedCrossRefGoogle Scholar
  20. 20.
    Plante M et al (1994) Interleukin-6 level in serum and ascites as a prognostic factor in patients with epithelial ovarian cancer. Cancer 73(7):1882–1888PubMedCrossRefGoogle Scholar
  21. 21.
    Scambia G et al (1995) Prognostic significance of interleukin 6 serum levels in patients with ovarian cancer. Br J Cancer 71(2):354–356PubMedCrossRefGoogle Scholar
  22. 22.
    Ishikawa H et al (2006) Mitogenic signals initiated via interleukin-6 receptor complexes in cooperation with other transmembrane molecules in myelomas. J Clin Exp Hematop 46(2):55–66PubMedCrossRefGoogle Scholar
  23. 23.
    Kanazawa T et al (2007) Interleukin-6 directly influences proliferation and invasion potential of head and neck cancer cells. Eur Arch Otorhinolaryngol 264(7):815–821PubMedCrossRefGoogle Scholar
  24. 24.
    Kenny HA et al (2008) The initial steps of ovarian cancer cell metastasis are mediated by MMP-2 cleavage of vitronectin and fibronectin. J Clin Invest 118(4):1367–1379PubMedCrossRefGoogle Scholar
  25. 25.
    Sawada K et al (2008) Loss of E-cadherin promotes ovarian cancer metastasis via alpha 5-integrin, which is a therapeutic target. Cancer Res 68(7):2329–2339PubMedCrossRefGoogle Scholar
  26. 26.
    Slack-Davis JK et al (2009) Vascular cell adhesion molecule-1 is a regulator of ovarian cancer peritoneal metastasis. Cancer Res 69(4):1469–1476PubMedCrossRefGoogle Scholar
  27. 27.
    Butler M et al (2007) Altered expression and endocytic function of CD205 in human dendritic cells, and detection of a CD205-DCL-1 fusion protein upon dendritic cell maturation. Immunology 120(3):362–371PubMedCrossRefGoogle Scholar
  28. 28.
    Zhu YX et al (2004) The SH3-SAM adaptor HACS1 is up-regulated in B cell activation signaling cascades. J Exp Med 200(6):737–747PubMedCrossRefGoogle Scholar
  29. 29.
    Mahnke K et al (2000) The dendritic cell receptor for endocytosis, DEC-205, can recycle and enhance antigen presentation via major histocompatibility complex class II-positive lysosomal compartments. J Cell Biol 151(3):673–684PubMedCrossRefGoogle Scholar
  30. 30.
    Wu K, Yuan J, Lasky LA (1996) Characterization of a novel member of the macrophage mannose receptor type C lectin family. J Biol Chem 271(35):21323–21330PubMedCrossRefGoogle Scholar
  31. 31.
    East L, Isacke CM (2002) The mannose receptor family. Biochim Biophys Acta 1572(2–3):364–386PubMedGoogle Scholar
  32. 32.
    Hawiger D et al (2001) Dendritic cells induce peripheral T cell unresponsiveness under steady state conditions in vivo. J Exp Med 194(6):769–779PubMedCrossRefGoogle Scholar
  33. 33.
    Bonifaz L et al (2002) Efficient targeting of protein antigen to the dendritic cell receptor DEC-205 in the steady state leads to antigen presentation on major histocompatibility complex class I products and peripheral CD8+ T cell tolerance. J Exp Med 196(12):1627–1638PubMedCrossRefGoogle Scholar
  34. 34.
    Trumpfheller C et al (2008) The microbial mimic poly IC induces durable and protective CD4+ T cell immunity together with a dendritic cell targeted vaccine. Proc Natl Acad Sci USA 105(7):2574–2579PubMedCrossRefGoogle Scholar
  35. 35.
    Kato M et al (2003) Hodgkin’s lymphoma cell lines express a fusion protein encoded by intergenically spliced mRNA for the multilectin receptor DEC-205 (CD205) and a novel C-type lectin receptor DCL-1. J Biol Chem 278(36):34035–34041PubMedCrossRefGoogle Scholar
  36. 36.
    Kato M et al (2007) The novel endocytic and phagocytic C-Type lectin receptor DCL-1/CD302 on macrophages is colocalized with F-actin, suggesting a role in cell adhesion and migration. J Immunol 179(9):6052–6063PubMedGoogle Scholar
  37. 37.
    al-Tubuly AA et al (1997) Inhibition of growth and enhancement of differentiation of colorectal carcinoma cell lines by MAb MR6 and IL-4. Int J Cancer 71(4):605–611PubMedCrossRefGoogle Scholar
  38. 38.
    Chapman EJ, Kelly G, Knowles MA (2008) Genes involved in differentiation, stem cell renewal, and tumorigenesis are modulated in telomerase-immortalized human urothelial cells. Mol Cancer Res 6(7):1154–1168PubMedCrossRefGoogle Scholar
  39. 39.
    Huarte E et al (2008) Depletion of dendritic cells delays ovarian cancer progression by boosting antitumor immunity. Cancer Res 68(18):7684–7691PubMedCrossRefGoogle Scholar

Copyright information

© Springer Science+Business Media B.V. 2011

Authors and Affiliations

  • Premkumar Vummidi Giridhar
    • 1
  • Holly M. Funk
    • 1
  • Catherine A. Gallo
    • 1
  • Aleksey Porollo
    • 2
  • Carol A. Mercer
    • 3
  • David R. Plas
    • 1
  • Angela F. Drew
    • 1
  1. 1.Department of Cancer and Cell Biology, Vontz Center for Molecular StudiesUniversity of CincinnatiCincinnatiUSA
  2. 2.Department of Environmental HealthUniversity of CincinnatiCincinnatiUSA
  3. 3.PDS BiotechnologyLawrenceburgUSA

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