Abstract
In prostate cancer metastases to bone, cancer cell-derived cytokines stimulate RANKL expression by cells of the osteoblast lineage, which in turn activates osteoclastic bone resorption. However, it is unclear whether cells of the osteoblast lineage signal back to prostate cancer cells, and if so, whether such direct cross-talk can be targeted therapeutically. Using the human prostate cancer cell line, PC3, we identified two novel signalling pathways acting between cells of the osteoblast lineage and cancer cells. First, exposure to RANKL stimulated the expression and release of IL-6 by PC3 cells in vitro (which is known to promote RANKL expression by osteoblasts). Second, treatment of PC3 cells with IL-6 increased the expression of RANK, the cognate receptor of RANKL, and enhanced the RANKL-induced release of IL-6 by PC3 cells. Third, targeted disruption of IL-6 signaling with tocilizumab, a clinically available antibody against the human IL-6 receptor, inhibited skeletal tumor growth in vivo and reduced serum RANKL levels as well as RANK expression by PC3-derived bone tumors. Similar effects were achieved when RANK expression was knocked down in PC3 cells. In contrast, disruption of IL-6 or RANK/RANKL signalling had no effect on PC3 tumor growth in soft tissues, indicating that these signalling pathways act specifically within the bone microenvironment. In conclusion, prostate cancer cells and cells of the osteoblast lineage communicate via two inter-dependent signaling pathways, which through auto-amplification strongly enhance metastatic prostate cancer growth in bone. Both pathways may be targeted for effective therapeutic intervention.
Similar content being viewed by others
References
Coleman RE (2001) Metastatic bone disease: clinical features, pathophysiology and treatment strategies. Cancer Treat Rev 27(3):165–176
Weilbaecher KN, Guise TA, McCauley LK (2011) Cancer to bone: a fatal attraction. Nat Rev Cancer 11(6):411–425
Zheng Y, Zhou H, Brennan K, Blair JM, Modzelewski JR, Seibel MJ, Dunstan CR (2007) Inhibition of bone resorption, rather than direct cytotoxicity, mediates the anti-tumour actions of ibandronate and osteoprotegerin in a murine model of breast cancer bone metastasis. Bone 40(2):471–478
Neudert M, Fischer C, Krempien B, Bauss F, Seibel MJ (2003) Site-specific human breast cancer (MDA-MB-231) metastases in nude rats: model characterisation and in vivo effects of ibandronate on tumour growth. Int J Cancer 107(3):468–477
Armstrong AP, Miller RE, Jones JC, Zhang J, Keller ET, Dougall WC (2008) RANKL acts directly on RANK-expressing prostate tumor cells and mediates migration and expression of tumor metastasis genes. Prostate 68(1):92–104
Price JT, Quinn JM, Sims NA, Vieusseux J, Waldeck K, Docherty SE, Myers D, Nakamura A, Waltham MC, Gillespie MT, Thompson EW (2005) The heat shock protein 90 inhibitor, 17-allylamino-17-demethoxygeldanamycin, enhances osteoclast formation and potentiates bone metastasis of a human breast cancer cell line. Cancer Res 65(11):4929–4938
Zheng Y, Zhou H, Ooi LL, Snir AD, Dunstan CR, Seibel MJ (2011) Vitamin D deficiency promotes prostate cancer growth in bone. Prostate 71(9):1012–1021
Ooi LL, Zheng Y, Zhou H, Trivedi T, Conigrave AD, Seibel MJ, Dunstan CR (2010) Vitamin D deficiency promotes growth of MCF-7 human breast cancer in a rodent model of osteosclerotic bone metastasis. Bone 47(4):795–803
Mundy GR (1997) Mechanisms of bone metastasis. Cancer 80(Suppl 8):1546–1556
Guise TA (2002) The vicious cycle of bone metastases. J Musculoskelet Neuronal Interact 2(6):570–572
Kamiya N, Suzuki H, Yano M, Endo T, Takano M, Komaru A, Kawamura K, Sekita N, Imamoto T, Ichikawa T (2010) Implications of serum bone turnover markers in prostate cancer patients with bone metastasis. Urology 75(6):1446–1451
Michalaki V, Syrigos K, Charles P, Waxman J (2004) Serum levels of IL-6 and TNF-alpha correlate with clinicopathological features and patient survival in patients with prostate cancer. Br J Cancer 90(12):2312–2316
George DJ, Halabi S, Shepard TF, Sanford B, Vogelzang NJ, Small EJ, Kantoff PW (2005) The prognostic significance of plasma interleukin-6 levels in patients with metastatic hormone-refractory prostate cancer: results from cancer and leukemia group B 9480. Clin Cancer Res 11(5):1815–1820
Wegiel B, Bjartell A, Culig Z, Persson JL (2008) Interleukin-6 activates PI3 K/Akt pathway and regulates cyclin A1 to promote prostate cancer cell survival. Int J Cancer 122(7):1521–1529
Domingo-Domenech J, Oliva C, Rovira A, Codony-Servat J, Bosch M, Filella X, Montagut C, Tapia M, Campas C, Dang L, Rolfe M, Ross JS, Gascon P, Albanell J, Mellado B (2006) Interleukin 6, a nuclear factor-kappaB target, predicts resistance to docetaxel in hormone-independent prostate cancer and nuclear factor-kappaB inhibition by PS-1145 enhances docetaxel antitumor activity. Clin Cancer Res 12(18):5578–5586
Hong DS, Angelo LS, Kurzrock R (2007) Interleukin-6 and its receptor in cancer: implications for Translational Therapeutics. Cancer 110(9):1911–1928
De La Mata J, Uy HL, Guise TA, Story B, Boyce BF, Mundy GR, Roodman GD (1995) Interleukin-6 enhances hypercalcemia and bone resorption mediated by parathyroid hormone-related protein in vivo. J Clin Invest 95(6):2846–2852
Suda T, Takahashi N, Udagawa N, Jimi E, Gillespie MT, Martin TJ (1999) Modulation of osteoclast differentiation and function by the new members of the tumor necrosis factor receptor and ligand families. Endocr Rev 20(3):345–357
Franchimont N, Lambert C, Huynen P, Ribbens C, Relic B, Chariot A, Bours V, Piette J, Merville MP, Malaise M (2005) Interleukin-6 receptor shedding is enhanced by interleukin-1beta and tumor necrosis factor alpha and is partially mediated by tumor necrosis factor alpha-converting enzyme in osteoblast-like cells. Arthritis Rheum 52(1):84–93
Kudo O, Sabokbar A, Pocock A, Itonaga I, Fujikawa Y, Athanasou NA (2003) Interleukin-6 and interleukin-11 support human osteoclast formation by a RANKL-independent mechanism. Bone 32(1):1–7
Sansone P, Storci G, Tavolari S, Guarnieri T, Giovannini C, Taffurelli M, Ceccarelli C, Santini D, Paterini P, Marcu KB, Chieco P, Bonafe M (2007) IL-6 triggers malignant features in mammospheres from human ductal breast carcinoma and normal mammary gland. J Clin Invest 117(12):3988–4002
Schafer ZT, Brugge JS (2007) IL-6 involvement in epithelial cancers. J Clin Invest 117(12):3660–3663
Kim MY, Oskarsson T, Acharyya S, Nguyen DX, Zhang XH, Norton L, Massague J (2009) Tumor self-seeding by circulating cancer cells. Cell 139(7):1315–1326
Sohara Y, Shimada H, Minkin C, Erdreich-Epstein A, Nolta JA, DeClerck YA (2005) Bone marrow mesenchymal stem cells provide an alternate pathway of osteoclast activation and bone destruction by cancer cells. Cancer Res 65(4):1129–1135
Ara T, Song L, Shimada H, Keshelava N, Russell HV, Metelitsa LS, Groshen SG, Seeger RC, DeClerck YA (2009) Interleukin-6 in the bone marrow microenvironment promotes the growth and survival of neuroblastoma cells. Cancer Res 69(1):329–337
Bendre MS, Margulies AG, Walser B, Akel NS, Bhattacharrya S, Skinner RA, Swain F, Ramani V, Mohammad KS, Wessner LL, Martinez A, Guise TA, Chirgwin JM, Gaddy D, Suva LJ (2005) Tumor-derived interleukin-8 stimulates osteolysis independent of the receptor activator of nuclear factor-kappaB ligand pathway. Cancer Res 65(23):11001–11009
Bussard KM, Venzon DJ, Mastro AM (2010) Osteoblasts are a major source of inflammatory cytokines in the tumor microenvironment of bone metastatic breast cancer. J Cell Biochem 111(5):1138–1148
Paule B, Terry S, Kheuang L, Soyeux P, Vacherot F, de la Taille A (2007) The NF-kappaB/IL-6 pathway in metastatic androgen-independent prostate cancer: new therapeutic approaches? World J Urol 25(5):477–489
Santini D, Perrone G, Roato I, Godio L, Pantano F, Grasso D, Russo A, Vincenzi B, Fratto ME, Sabbatini R, Della Pepa C, Porta C, Del Conte A, Schiavon G, Berruti A, Tomasino RM, Papotti M, Papapietro N, Muda AO, Denaro V, Tonini G (2011) Expression pattern of receptor activator of NFκB (RANK) in a series of primary solid tumors and related bone metastases. J Cell Physiol 226(3):780–784
Jones DH, Nakashima T, Sanchez OH, Kozieradzki I, Komarova SV, Sarosi I, Morony S, Rubin E, Sarao R, Hojilla CV, Komnenovic V, Kong YY, Schreiber M, Dixon SJ, Sims SM, Khokha R, Wada T, Penninger JM (2006) Regulation of cancer cell migration and bone metastasis by RANKL. Nature 440(7084):692–696
Xiong J, Onal M, Jilka RL, Weinstein RS, Manolagas SC, O’Brien CA (2011) Matrix-embedded cells control osteoclast formation. Nat Med 17(10):1235–1241
Zheng Y, Zhou H, Fong-Yee C, Modzelewski JR, Seibel MJ, Dunstan CR (2008) Bone resorption increases tumour growth in a mouse model of osteosclerotic breast cancer metastasis. Clin Exp Metastasis 25(5):559–567
Zheng Y, Seibel M, Zhou H (2011) Methods in bone biology: cancer and bone. Osteoporosis research: animal models, 1st edn. Springer, New York
Lacey DL, Timms E, Tan HL, Kelley MJ, Dunstan CR, Burgess T, Elliott R, Colombero A, Elliott G, Scully S, Hsu H, Sullivan J, Hawkins N, Davy E, Capparelli C, Eli A, Qian YX, Kaufman S, Sarosi I, Shalhoub V, Senaldi G, Guo J, Delaney J, Boyle WJ (1998) Osteoprotegerin ligand is a cytokine that regulates osteoclast differentiation and activation. Cell 93(2):165–176
Ehlers M, Grotzinger J, deHon FD, Mullberg J, Brakenhoff JP, Liu J, Wollmer A, Rose-John S (1994) Identification of two novel regions of human IL-6 responsible for receptor binding and signal transduction. J Immunol 153(4):1744–1753
Igawa T, Ishii S, Tachibana T, Maeda A, Higuchi Y, Shimaoka S, Moriyama C, Watanabe T, Takubo R, Doi Y, Wakabayashi T, Hayasaka A, Kadono S, Miyazaki T, Haraya K, Sekimori Y, Kojima T, Nabuchi Y, Aso Y, Kawabe Y, Hattori K (2011) Antibody recycling by engineered pH-dependent antigen binding improves the duration of antigen neutralization. Nat Biotechnol 28(11):1203–1207
Axmann R, Böhm C, Krönke G, Zwerina J, Smolen J, Schett G (2009) Inhibition of interleukin-6 receptor directly blocks osteoclast formation in vitro and in vivo. Arthritis Rheum 60(9):2747–2756
Blair JM, Zheng Y, Dunstan CR (2007) RANK ligand. Int J Biochem Cell Biol 39(6):1077–1081
Takayanagi H, Ogasawara K, Hida S, Chiba T, Murata S, Sato K, Takaoka A, Yokochi T, Oda H, Tanaka K, Nakamura K, Taniguchi T (2000) T-cell-mediated regulation of osteoclastogenesis by signalling cross-talk between RANKL and IFN-[gamma]. Nature 408(6812):600–605
Shariat SF, Andrews B, Kattan MW, Kim J, Wheeler TM, Slawin KM (2001) Plasma levels of interleukin-6 and its soluble receptor are associated with prostate cancer progression and metastasis. Urology 58(6):1008–1015
Salgado R, Junius S, Benoy I, Van Dam P, Vermeulen P, Van Marck E, Huget P, Dirix LY (2003) Circulating interleukin-6 predicts survival in patients with metastatic breast cancer. Int J Cancer 103(5):642–646
Ignatoski KM, Friedman J, Escara-Wilke J, Zhang X, Daignault S, Dunn RL, Smith DC, Keller ET (2009) Change in markers of bone metabolism with chemotherapy for advanced prostate cancer: interleukin-6 response is a potential early indicator of response to therapy. J Interferon Cytokine Res 29(2):105–112
Nishimoto N, Kishimoto T (2008) Humanized antihuman IL-6 receptor antibody, tocilizumab. Handb Exp Pharmacol 181:151–160
Shinriki S, Jono H, Ota K, Ueda M, Kudo M, Ota T, Oike Y, Endo M, Ibusuki M, Hiraki A, Nakayama H, Yoshitake Y, Shinohara M, Ando Y (2009) Humanized Anti-Interleukin-6 Receptor Antibody Suppresses Tumor Angiogenesis and In vivo Growth of Human Oral Squamous Cell Carcinoma. Clin Cancer Res 15(17):5426–5434
Sakai A, Oda M, Itagaki M, Yoshida N, Arihiro K, Kimura A (2010) Establishment of an HS23 stromal cell-dependent myeloma cell line: fibronectin and IL-6 are critical. Int J Hematol 92(4):598–608
Guise TA, Mohammad KS, Clines G, Stebbins EG, Wong DH, Higgins LS, Vessella R, Corey E, Padalecki S, Suva L, Chirgwin JM (2006) Basic Mechanisms Responsible for Osteolytic and Osteoblastic Bone Metastases. Clin Cancer Res 12(20):6213s–6216s
Ooi LL, Zheng Y, Stalgis-Bilinski K, Dunstan CR (2011) The bone remodeling environment is a factor in breast cancer bone metastasis. Bone 48(1):66–70
Zheng Y, Zhou H, Dunstan C, Sutherland RL, Seibel M (2013) The role of the bone microenvironment in skeletal metastasis. J Bone Oncol 2(1):47–57
Guise TA, Yin JJ, Thomas RJ, Dallas M, Cui Y, Gillespie MT (2002) Parathyroid hormone-related protein (PTHrP)-(1-139) isoform is efficiently secreted in vitro and enhances breast cancer metastasis to bone in vivo. Bone 30(5):670–676
Zhang J, Dai J, Qi Y, Lin DL, Smith P, Strayhorn C, Mizokami A, Fu Z, Westman J, Keller ET (2001) Osteoprotegerin inhibits prostate cancer-induced osteoclastogenesis and prevents prostate tumor growth in the bone. J Clin Invest 107(10):1235–1244
Brown JE, Cook RJ, Major P, Lipton A, Saad F, Smith M, Lee KA, Zheng M, Hei YJ, Coleman RE (2005) Bone turnover markers as predictors of skeletal complications in prostate cancer, lung cancer, and other solid tumors. J Natl Cancer Inst 97(1):59–69
Berruti A, Tucci M, Mosca A, Tarabuzzi R, Gorzegno G, Terrone C, Vana F, Lamanna G, Tampellini M, Porpiglia F, Angeli A, Scarpa RM, Dogliotti L (2005) Predictive factors for skeletal complications in hormone-refractory prostate cancer patients with metastatic bone disease. Br J Cancer 93(6):633–638
Fizazi K, Carducci M, Smith M, Damiao R, Brown J, Karsh L, Milecki P, Shore N, Rader M, Wang H, Jiang Q, Tadros S, Dansey R, Goessl C (2011) Denosumab versus zoledronic acid for treatment of bone metastases in men with castration-resistant prostate cancer: a randomised, double-blind study. Lancet 377(9768):813–822
Fizazi K, Bosserman L, Gao G, Skacel T, Markus R (2013) Denosumab treatment of prostate cancer with bone metastases and increased urine N-telopeptide levels after therapy with intravenous bisphosphonates: results of a randomized phase II trial. J Urol 189(Suppl 1):S51–S58
Coleman R, Gnant M, Morgan G, Clezardin P (2012) Effects of bone-targeted agents on cancer progression and mortality. J Natl Cancer Inst 104(14):1059–1067
Smith MR, Saad F, Coleman R, Shore N, Fizazi K, Tombal B, Miller K, Sieber P, Karsh L, Damiao R, Tammela TL, Egerdie B, Van Poppel H, Chin J, Morote J, Gomez-Veiga F, Borkowski T, Ye Z, Kupic A, Dansey R, Goessl C (2012) Denosumab and bone-metastasis-free survival in men with castration-resistant prostate cancer: results of a phase 3, randomised, placebo-controlled trial. Lancet 379(9810):39–46
Acknowledgments
This work has been supported in part by the following Grants: Cure Cancer Foundation of Australia (Y.Z), Cancer Institute New South Wales (Y.Z), Cancer Council New South Wales (M.J.S, Y.Z, H.Z, C.R.D), Prostate Cancer Foundation of Australia (PCFA) (M.J.S, Y.Z, H.Z, C.R.D), University of Sydney Cancer Research Fund (M.J.S), National Health and Medical Research Council, Australia (NHMRC) (Y.Z, Early Career Fellowship 596870) and Bundesministerium für Bildung und Forschung (BMBF) and the State of Berlin (K.B. F.B., BCRT-Grant I and II). We thank Dr. Matthew Foley and the staff at the Australian Centre for Microscopy & Microanalysis, The University of Sydney for the facilities as well as scientific and technical assistance.
Conflict of interest
The authors disclose no potential conflicts of interest.
Author information
Authors and Affiliations
Corresponding authors
Rights and permissions
About this article
Cite this article
Zheng, Y., Basel, D., Chow, SO. et al. Targeting IL-6 and RANKL signaling inhibits prostate cancer growth in bone. Clin Exp Metastasis 31, 921–933 (2014). https://doi.org/10.1007/s10585-014-9680-3
Received:
Accepted:
Published:
Issue Date:
DOI: https://doi.org/10.1007/s10585-014-9680-3