Tumor and Host Determinants of Pulmonary Metastasis in Bladder Cancer

  • Neveen Said
  • Dan Theodorescu


Despite the recent advances in the diagnosis of bladder cancer, ­recurrence after surgical intervention for muscle invasive disease is still problematic as nearly half of patients harbor occult distant metastases. Clinical data from human disease revealed that, invasive and metastatic bladder cancer cells can metastasize to lungs, and this in turn is associated with poor 5-year survival rate. Experimental rodent models of carcinogenesis and metastasis are available to study this phenomenon. Comparative gene expression profiling, proteomic and computational studies identified an intertwined network of metastasis promoters and suppressors that modulate the interactions between the components of the pulmonary milieu and cancer cells inflammatory mediators, ECM molecules, as well as peptide hormones. In this chapter we provide select exemplar of some of the molecular mechanisms underlying lung colonization by bladder cancer.


Bladder Cancer Bladder Cancer Cell Urothelial Cancer Human Bladder Cancer Bladder Cancer Cell Line 
These keywords were added by machine and not by the authors. This process is experimental and the keywords may be updated as the learning algorithm improves.

14.1 Introduction

Urothelial cancer (UC) is the most common malignancy affecting the urinary ­system. When it affects the bladder, it leads to an estimated 70,530 new cases in the United States, with a male to female ratio of 3:1 and approximately 14,680 deaths, expected in 2010 [1]. UC arises from the mucosal lining and is frequently ­multifocal. Numerous factors, including chromosomal markers, genetic ­polymorphisms, and genetic and epigenetic alterations may be involved in tumorigenesis, progression and metastasis. At initial presentation, 70% of patients with UC present with non-muscle invasive (formerly known as “superficial”), and 30% present with muscle invasive disease [2]. Despite a good prognosis for patients with the former, recurrence is common and is associated with development of muscle invasive disease in up to 30%. Also, 50% of patients presenting with muscle invasive UC develop distant metastases leading to poor 5-year survival rate [1]. Clinical data from human disease as well as experimental rodent models of carcinogenesis and metastasis reveals that when it occurs, metastasis of this tumor is commonly found in the regional lymph node metastasis and the lungs. Given that metastases are responsible for most of the deaths from this disease [3, 4], understanding of the process will aid in the development of new approaches for treatment.

14.2 Multistep Process of Metastasis

The high incidence of pulmonary metastases in cancer patients was initially believed to be a random process based on blood flow predominance as lungs receive significant cardiac output [5, 6]. This was supported by the observation that metastases often initiate in pulmonary arterioles and later traverse the basement membrane into the lung parenchyma [6, 7]. However, growing body of literature supports the patterns of metastases is also a consequence of the “seed and soil” theory put forth by Stephen Paget in 1889 [8]. Hence, the development of lung metastasis is likely an active highly selective process instigated by tumor cells, and is strongly influenced by the interactions between tumor and host cells, and by both the immediate and extended tumor microenvironments [3, 7, 8, 9, 10, 11, 12, 13, 14].

14.2.1 The Lung Microenvironment as a Host for Metastatic Tumor Cells

Barriers created by the lung microenvironment include physical barriers such as vascular endothelium, extracellular matrix (ECM) components and basement ­membranes, as well as physiologic barriers such as limited oxygen (hypoxia) and nutrients, and immunologic barriers by the immune system [9, 15]. The vascular endothelium is a critical regulator of pulmonary function and its phenotype is modulated by myriad stimuli such as proinflammatory cytokines and hemodynamic forces thereby impacting the development of vascular disease states [15]. Metastatic progression also depends on cancer cell secretome as both the growing primary tumor and shed tumor cells in the circulation (Fig. 14.1), secrete ­inflammatory mediators that triggers vascular inflammation in the lungs that is characterized by recruitment of bone marrow derived myeloid cells, margination, extravasation, and activation of circulating mononuclear cells into vessel walls to lung parenchyma [10, 12, 13, 14, 16, 17, 18, 19]. This multistep process is the result of local and systemic secretion of chemotactic factors, macrophage-activating cytokines, endothelin-1, reactive oxygen species (ROS), growth factors, upregulation of vascular adhesion molecules, and production of MMPs. The buildup of an inflammatory microenvironment, generates a premetastatic niche with progressive pulmonary vascular inflammation hospitable for metastatic growth [10, 12, 13, 14, 18, 19]. The activation of endothelial cells (ECs) is a crucial step in pulmonary vascular inflammation, and can result in subsequent increase in surface expression of cell adhesion molecules, such as vascular cell adhesion molecule (VCAM)-1, intercellular adhesion ­molecule (ICAM)-1, and selectins, which contribute to the recruitment of inflammatory cells to ECs and their transmigration across the vascular wall [18, 20]. In turn, activated ECs secrete cytokines and chemokines such as monocyte chemotactic protein 1 (MCP-1), which is a potent inducer for ­monocyte attachment to ECs and ­migration into subendothelial space [21, 22, 23, 24]. Mechanisms leading to proinflammatory cell recruitment to the lung are exploited by circulating cancer cells to extravasate from circulation [20]. These mechanisms include common cell adhesion molecule interactions as well as the expression of inflammatory cytokines/chemokines leading to recruitment and activation of macrophages. Persistent activation of ECs and macrophages, with reciprocal persistent production of cytokines and ROS cause local chronic inflammation that promotes increased homing of tumor cells to the lungs [21, 22, 23, 24] as well as activation of non-tumor cells in tumor lesions stimulating tumor angiogenesis, invasiveness and intravasation/extravasation. Interestingly, a strong body of literature highlighted the central importance of nuclear factor-κB (NF-κB) in the activation of and cross-talk between all the cell types involved [22, 23, 24].
Fig. 14.1

A view of the multistep process of metastasis: Disseminated cancer cells in the circulation arise from highly invasive cell populations in the primary tumor that invade the blood vessel wall, intravasate and circulate in the blood stream. Secreted tumor factors “cancer secretome” form invasive cancer cells in the primary tumors and circulating tumor cells recruit, bone marrow-derived myeloid cells, to the prospective metastatic site “lungs”. In turn, myeloid cells acquire an inflammatory macrophage phenotype, secreting factors “macrophage secretome” and contribute to the construction of a pre-metastatic niche favorable for implantation and growth of disseminating tumor cells forming micro- and macro-metastases

The premetastatic niche may also be responsible for metastases to specific organs. The key tumor-secreted factors that determine metastatic sites and mediate premetastatic niche formation have yet to be identified, although the roles for cytokines/chemokines, MMPs and growth factors’ signaling in pulmonary endothelial cells and macrophages have been reported [25, 26]. These make the lung microenvironment more receptive to cancers cells. We have recently reported the roles of endothelin-1 in as an important mediator of lung inflammation and colonization of bladder cancer cells (discussed below) [14]. Elevated fibronectin expression by fibroblasts and fibroblast-like cells resident at premetastatic sites seems to be an important factor in the development of the premetastatic niche [12]. In addition, the secretion of versican by tumor cells and subsequently by inflammatory macrophages has been recently found to play a role in lung metastasis [10, 16, 27].

Overexpression of tumor-derived immunosuppressive factors such as TGFβ, VEGF, IL-6, MCP1/CCL2, MMPs and Cox-2/prostanoids in the inflammatory premetastatic niche dampens the tumor-suppressing activities of cells responsible of immune surveillance as natural killer cells and antigen-presenting cells and supports an inflammatory phenotype of tumor-associated macrophages, TAM; promote tumor progression by secreting growth factors and cytokines that stimulate tumor metastases [16, 17, 28]. TAMs can facilitate tumor formation and progression by activation of NF-κB and AP-1 [16, 17, 29], and are especially attracted to regions of hypoxia, where they secrete angiogenic inducers and proteases, and express high levels of the HIF-1 and -2 transcription factors [18, 30, 31, 32] that further augment macrophage recruitment and activation as well as tumor cell invasiveness. Furthermore, TAMs can release growth factors such as PDGF, EGF, cytokines/chemokines, and MMPs which enhance proliferation, survival, and invasion of tumor cells as well as secrete and induce secretion of ECM proteoglycans such as versican from tumor cells and fibroblasts in the lung milieu further augmenting invasiveness, inflammation and metastasis [10, 23, 33, 34, 35, 36, 37, 38].

Our laboratory has identified two major molecules implicated in bladder cancer metastasis to the lungs, the metastasis suppressor RhoGDI2 and the pro-metastatic Ral A/B.

14.2.2 The Metastasis Suppressor RhoGDI2

Rho family GDP Dissociation Inhibitor 2 (RhoGDI2) protein has been identified as functional metastasis suppressor during studies of the differential invasive and metastatic properties of isogenic human bladder carcinoma cell lines, T24 (non-­metastatic), and T24T (highly invasive and metastatic) using experimental metastasis models and comparative genomic studies [39]. RhoGDI2 was also found to be a prognostic marker in patients after cystectomy that its reduced expression was associated with decreased patient survival. However, approximately 35% of patients with moderate or high levels of RhoGDI2 protein-developed metastatic disease ­suggesting that not only the expression level, but other mechanisms might regulate the metastasis suppressor effect of RhoGDI2 [40]. Phosphorylation, binding to specific partners, truncations, proteolytic cleavage, or change in subcellular localization was considered given the existing data on the other members of the RhoGDI family (reviewed in [41]). Comparative gene expression profiling of human bladder cancer tissues and cell lines identified that the down-stream effectors downregulated by RhoGDI2 are molecules that have long been implicated in pulmonary vascular diseases: Endothelin-1 (ET-1) [42] and versican [14, 27, 43].

14.2.3 Endothelin Axis

ET-1, an endothelial cell-derived vasoconstrictor peptide is an important member of the endothelin axis with myriad developmental, physiological and pathological functions [29, 44, 45, 46]. The “Endothelin Axis” consists of three similar small peptides, ET-1, ET-2 and ET-3, two G-protein-coupled receptors, ETAR and ETBR, and two membrane-bound proteases, the ET-converting enzymes, ECE-1 and ECE-2 [44], that activate the secreted pro forms of the peptide. ET-1 production is stimulated by a variety of cytokines and growth factors, hypoxia, and shear stress, while ETAR activation triggers signaling networks involved in cell proliferation, new vessel formation, invasion, inflammation, and metastatic spread [44, 47, 48, 49]. In benign and malignant diseases, ET-1 has been shown to activate the pro-inflammatory transcriptional factors AP-1 and NF-κB in human monocytes and cancer cells (Fig. 14.2) and to stimulate the production of inflammatory cytokines IL-6, MCP-1 and Cox2, as well as MMP activity, the key orchestrators of inflammation-mediated cancer cell invasiveness and metastasis [50, 51, 52, 53]. Recently, gene expression data and immunostaining of from human bladder cancer has revealed that ET-1 and ETAR are over-expressed in muscle invasive disease, and that the expression level is associated with reduced patient survival [14]. Experimental and spontaneous metastasis models revealed that tumor ET-1 triggers inflammation in the lungs wherein inflammatory cells, primarily macrophages, enhance and facilitate the process of metastatic colonization [14]. The magnitude of this early inflammatory response determines the kinetics of subsequent development of lung metastases. This inflammatory response is likely triggered by tumor secretion of ET-1 and the pro-inflammatory cytokines IL-6 and MCP-1. Interestingly, targeting ET-1 gene in tumor cells or pharmacologic inhibition of ETAR prior to injection of tumor cells reduced the early inflammatory response, both tumor and host MCP-1/CCL2, IL-6 and Cox-2 in the lung microenvironment and subsequent metastatic colonization. In contrast, when the early inflammatory response was allowed to develop upon injection of tumor cells, then later followed by ETAR blockade, the reduction of lung inflammation and clinical lung metastases was not as dramatic. Depletion of macrophages also significantly reduced the inflammatory response and the ensuing lung metastases.
Fig. 14.2

Multiple pathways of endothelin involvement in lung metastasis: (a) Binding of ET-1 to the ETA receptor in the plasma membrane triggers signal-transduction pathways in both cancer cells and macrophages that converge in activation of NF-κB and AP-1 transcription factors. (b) Schematic illustration of the roles of ET-1 in bladder cancer metastasis to the lungs

14.2.4 Versican

Versican, a chondroitin sulfate proteoglycan, is a major structural component of the ECM (Fig. 14.3). Differential RNA splicing gives rise to four isoforms of versican (V0, V1, V2, and V3), which vary by the presence or absence of two glycosaminoglycan (GAG) binding domains named αGAG and βGAG. The various domains of versican form highly hydrated complexes that trap cytokines, enzymes, growth factors, lipoproteins, other extracellular matrix molecules, and signaling receptors. It is implicated in the pathogenesis of pulmonary vascular diseases and atherosclerosis, causing an ECM expansion, fostering pro-inflammatory cytokines and cells and dramatically influencing the inflammatory phenotype of cellular components of these diseases [54].
Fig. 14.3

Structure of versican and its interactions: The N-terminal, G1 domain is composed of an immunoglobulin (Ig)-like motif, followed by two proteoglycan tandem repeats that bind to hyaluronan. The C-terminal, G3 domain, consists of two epidermal growth factor (EGF)-like repeats, a carbohydrate recognition domain (CRD), and a complement binding protein (CBP). Chondroitin sulfate (CS) chain binding regions exist between G1 and G3 consist of a protein core and one or two glycosaminoglycans (α and β GAG) that vary amongst tissues and specific molecules. Different domains of versican bind to and interact with a wide variety of molecules, such as growth factors (TGFβ, PDGF-BB), cytokines, chemokines (MCP-1), adhesion/ECM molecules (selectins, HA, CD44, fibrinogen/fibrin, fibronectin, thrombospondin, collagens), MMPs and LDL

The association between increased levels of versican and progression of cancer to disseminated disease suggests that versican is important in promoting cancer cell proliferation, motility, invasion and metastasis [10, 16, 23, 33, 34, 35, 36, 37, 38, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65]. More recent studies [10] identified cancer cell-secreted versican, as a macrophage activator that acts through TLR2/6 inducing TNF-alpha secretion by myeloid cells, generating an inflammatory microenvironment hospitable for metastatic growth. In vitro functional studies, revealed versican is a key mediator of cancer cell-macrophages cross-talk and significantly inhibited their adhesion to and invasion of ECM molecules as well as pulmonary microvascular endothelial cells [27, 66]. Interestingly, comparative gene expression profiling of invasive/metastatic bladder cancer cells with clinical bladder cancer specimens identified versican as the top upregulated gene, positively correlated with bladder cancer stage and poor prognosis, and in vitro with invasiveness of bladder cancer cell lines [27, 43]. It remains to be seen if genetic manipulation of versican expression in bladder cancer cells, either by stable overexpression or knockdown demonstrates a role of tumor versican in metastasis.

14.2.5 Ral GTPases

The Ras-like family of monomeric G proteins (Ral GTPases), RalA and RalB are paralogs (85% amino acid identity) that participate in diverse cellular functions [67] (Fig. 14.4). These signaling proteins, regulated by multiple pathways including Ras (reviewed in [68]), have been shown to be necessary for cellular phenotypes associated with cancer progression and metastasis. Phosphorylation and subsequent activation, of RalA and RalB is paralog-specific as they are phosphorylated at specific conserved sites by different kinases [69]. PKC phosphorylation of RalB at S198 in bladder cancer cell lines has been recently reported to be necessary for cytoskeletal organization, anchorage-independent growth, and cell migration in vitro and for subcutaneous tumor growth and lung metastasis in vivo [69]. In addition, Ral effector, RalBP1 was found to be highly expressed in bladder cancer patients and necessary for metastasis of human bladder cancer cell lines [70]. In addition RalA and/or RalB have also been shown to regulate transcription factors such as NF-κB and AP-1 [71, 72] and play a role in the regulation of expression of key molecules, including the prognostic marker and cell surface GPI-linked glycoprotein, CD24 [68].
Fig. 14.4

The regulation of Ral and downstream signaling: RalGTPases are activated by a variety of stimuli, including guanidine nucleotide exchange factors (RalGEFs), Ras or independent of Ras activation. Several other stimuli, known to be active in cancer cells, have been identified to activate Ral, including oxidative damage, hypoxia, calcium signaling, and androgen withdrawal. Once activated by exchange of GTP for GDP, Ral GTPases are able to associate physically with several known (and likely other uncharacterized) effector proteins, to mediate diverse cellular effects

14.2.6 CD24

CD24, is a glycosyl phosphatidyl inositol–linked surface protein [73], that has been identified as a downstream target of Ral signaling by profiling the expression of RalA/B–depleted bladder carcinoma cells [67, 68]. Originally identified as a surface differentiation marker of peripheral B-lymphocytes [73], CD24 received recent attention as a marker of stem cell populations of a variety of cancers [74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87]. CD24 overexpression has been associated with lung metastasis [88] as it promoted cancer cell rolling and adhesion to P-selectin in murine lung vasculature and platelets [76, 78, 79, 80, 85, 89, 90, 91, 92, 93, 94, 95]. Consistently, CD24 has been recently reported to be a hypoxia-regulated gene in some cancer cell lines [96, 97] that was down-regulated by Cox-2 inhibitor, Celecoxib, suggesting role in inflammation associated with metastasis [98, 99, 100]. CD24 is highly expressed in urothelial as well as other cancers. Immunohistological localization of CD24 revealed high levels of expression correlated with the stromal/muscle invasion, stage, grade and shorter patient disease-free survival [67, 101]. Depletion of CD24 function in bladder cancer cell lines was found to be associated with decreased cell proliferation and anchorage-independent growth, changes in the actin cytoskeleton, and induction of apoptosis [67]. In addition, monoclonal antibodies against CD24 that were efficacious in inhibiting in vitro and in vivo growth of colorectal and pancreatic cancer cells as well as sensitizing them to conventional adjuvant chemotherapeutics [98, 99, 100, 102, 103, 104], significantly reduced the number of lung metastases that developed after tail vein injection of the metastatic UMUC3 cell line in an experimental model of bladder cancer [105]. However, the exact mechanism of CD24 involvement in lung metastasis is still elusive.


  1. 1.
    Jemal A, Siegel R, Xu J, Ward E (2010) Cancer statistics, 2010. CA Cancer J Clin 60:277–300PubMedGoogle Scholar
  2. 2.
    Wu XR (2005) Urothelial tumorigenesis: a tale of divergent pathways. Nat Rev Cancer 5:713–725PubMedGoogle Scholar
  3. 3.
    Rocken M (2010) Early tumor dissemination, but late metastasis: insights into tumor ­dormancy. J Clin Invest 120:1800–1803PubMedGoogle Scholar
  4. 4.
    Steeg PS, Theodorescu D (2008) Metastasis: a therapeutic target for cancer. Nat Clin Pract Oncol 5:206–219PubMedGoogle Scholar
  5. 5.
    Ewing J (1928) A treatise on tumors. In: Ewing J (ed) Neoplastic disease, 3rd edn. WB Saunders, Philadelphia, pp 77–89Google Scholar
  6. 6.
    Zetter BR (1990) The cellular basis of site-specific tumor metastasis. N Engl J Med 322:605–612PubMedGoogle Scholar
  7. 7.
    Luo JL, Maeda S, Hsu LC, Yagita H, Karin M (2004) Inhibition of NF-kappaB in cancer cells converts inflammation- induced tumor growth mediated by TNFalpha to TRAIL-mediated tumor regression. Cancer Cell 6:297–305PubMedGoogle Scholar
  8. 8.
    Paget S (1989) The distribution of secondary growths in cancer of the breast. 1889. Cancer Metastasis Rev 8:98–101PubMedGoogle Scholar
  9. 9.
    Gupta GP, Massague J (2006) Cancer metastasis: building a framework. Cell 127:679–695PubMedGoogle Scholar
  10. 10.
    Kim S, Takahashi H, Lin WW, Descargues P, Grivennikov S, Kim Y et al (2009) Carcinoma-produced factors activate myeloid cells through TLR2 to stimulate metastasis. Nature 457:102–106PubMedGoogle Scholar
  11. 11.
    Luo JL, Tan W, Ricono JM, Korchynskyi O, Zhang M, Gonias SL et al (2007) Nuclear cytokine-activated IKKalpha controls prostate cancer metastasis by repressing Maspin. Nature 446:690–694PubMedGoogle Scholar
  12. 12.
    Kaplan RN, Rafii S, Lyden D (2006) Preparing the “soil”: the premetastatic niche. Cancer Res 66:11089–11093PubMedGoogle Scholar
  13. 13.
    Kaplan RN, Riba RD, Zacharoulis S, Bramley AH, Vincent L, Costa C et al (2005) VEGFR1-positive haematopoietic bone marrow progenitors initiate the pre-metastatic niche. Nature 438:820–827PubMedGoogle Scholar
  14. 14.
    Said N, Smith SC, Sanchez-Carbayo M, Theodorescu D (2011) Tumor Endothelin-1 contributes to macrophage infiltration and metastatic colonization of murine lungs by human cancer cells. J Clin Invest 121(1):132–147Google Scholar
  15. 15.
    SenBanerjee S, Lin Z, Atkins GB, Greif DM, Rao RM, Kumar A et al (2004) KLF2 is a novel transcriptional regulator of endothelial proinflammatory activation. J Exp Med 199:1305–1315PubMedGoogle Scholar
  16. 16.
    Mantovani A (2009) Cancer: inflaming metastasis. Nature 457:36–37PubMedGoogle Scholar
  17. 17.
    Bidard FC, Pierga JY, Vincent-Salomon A, Poupon MF (2008) A “class action” against the microenvironment: do cancer cells cooperate in metastasis? Cancer Metastasis Rev 27:5–10PubMedGoogle Scholar
  18. 18.
    Tieu BC, Lee C, Sun H, Lejeune W, Recinos A 3rd, Ju X et al (2009) An adventitial IL-6/MCP1 amplification loop accelerates macrophage-mediated vascular inflammation leading to aortic dissection in mice. J Clin Invest 119:3637–3651PubMedGoogle Scholar
  19. 19.
    Qian B, Deng Y, Im JH, Muschel RJ, Zou Y, Li J et al (2009) A distinct macrophage population mediates metastatic breast cancer cell extravasation, establishment and growth. PLoS One 4:e6562PubMedGoogle Scholar
  20. 20.
    Taranova AG, Maldonado D 3rd, Vachon CM, Jacobsen EA, Abdala-Valencia H, McGarry MP et al (2008) Allergic pulmonary inflammation promotes the recruitment of circulating tumor cells to the lung. Cancer Res 68:8582–8589PubMedGoogle Scholar
  21. 21.
    Qi Y, Liang J, She ZG, Cai Y, Wang J, Lei T et al (2010) MCP-induced protein 1 suppresses TNFalpha-induced VCAM-1 expression in human endothelial cells. FEBS Lett 584(14):3065–3072PubMedGoogle Scholar
  22. 22.
    Rollins BJ (1996) Monocyte chemoattractant protein 1: a potential regulator of monocyte recruitment in inflammatory disease. Mol Med Today 2:198–204PubMedGoogle Scholar
  23. 23.
    Hess S, Methe H, Kim JO, Edelman ER (2009) NF-kappaB activity in endothelial cells is modulated by cell substratum interactions and influences chemokine-mediated adhesion of natural killer cells. Cell Transplant 18:261–273PubMedGoogle Scholar
  24. 24.
    Collins T, Cybulsky MI (2001) NF-kappaB: pivotal mediator or innocent bystander in atherogenesis? J Clin Invest 107:255–264PubMedGoogle Scholar
  25. 25.
    Hiratsuka S, Nakamura K, Iwai S, Murakami M, Itoh T, Kijima H et al (2002) MMP9 induction by vascular endothelial growth factor receptor-1 is involved in lung-specific metastasis. Cancer Cell 2:289–300PubMedGoogle Scholar
  26. 26.
    Hiratsuka S, Watanabe A, Aburatani H, Maru Y (2006) Tumour-mediated upregulation of chemoattractants and recruitment of myeloid cells predetermines lung metastasis. Nat Cell Biol 8:1369–1375PubMedGoogle Scholar
  27. 27.
    Said N, and Theodorescu, D. Role of Versican in Bladder Cancer Metastasis to Lungs. AACR-Joint Tumor Microenvironment and Metastasis Research Society Meeting; 2010 September, 2010; Philadelphia, PA; 2010.Google Scholar
  28. 28.
    Joyce JA, Pollard JW (2009) Microenvironmental regulation of metastasis. Nat Rev Cancer 9:239–252PubMedGoogle Scholar
  29. 29.
    Hagemann T, Balkwill F, Lawrence T (2007) Inflammation and cancer: a double-edged sword. Cancer Cell 12:300–301PubMedGoogle Scholar
  30. 30.
    Corzo CA, Condamine T, Lu L, Cotter MJ, Youn JI, Cheng P et al (2010) HIF-1alpha regulates function and differentiation of myeloid-derived suppressor cells in the tumor microenvironment. J Exp Med 207:2439–2453PubMedGoogle Scholar
  31. 31.
    Imtiyaz HZ, Williams EP, Hickey MM, Patel SA, Durham AC, Yuan LJ et al (2010) Hypoxia-inducible factor 2alpha regulates macrophage function in mouse models of acute and tumor inflammation. J Clin Invest 120:2699–2714PubMedGoogle Scholar
  32. 32.
    Doedens AL, Stockmann C, Rubinstein MP, Liao D, Zhang N, DeNardo DG et al (2010) Macrophage expression of hypoxia-inducible factor-1 alpha suppresses T-cell function and promotes tumor progression. Cancer Res 70:7465–7475PubMedGoogle Scholar
  33. 33.
    Ricciardelli C, Sakko AJ, Ween MP, Russell DL, Horsfall DJ (2009) The biological role and regulation of versican levels in cancer. Cancer Metastasis Rev 28:233–245PubMedGoogle Scholar
  34. 34.
    Lee JH, Kim SH, Lee ES, Kim YS (2009) CD24 overexpression in cancer development and progression: a meta-analysis. Oncol Rep 22:1149–1156PubMedGoogle Scholar
  35. 35.
    Yee AJ, Akens M, Yang BL, Finkelstein J, Zheng PS, Deng Z et al (2007) The effect of versican G3 domain on local breast cancer invasiveness and bony metastasis. Breast Cancer Res 9:R47PubMedGoogle Scholar
  36. 36.
    Wang W, Xu GL, Jia WD, Ma JL, Li JS, Ge YS et al (2009) Ligation of TLR2 by versican: a link between inflammation and metastasis. Arch Med Res 40:321–323PubMedGoogle Scholar
  37. 37.
    Chiodoni C, Colombo MP, Sangaletti S (2010) Matricellular proteins: from homeostasis to inflammation, cancer, and metastasis. Cancer Metastasis Rev 29:295–307PubMedGoogle Scholar
  38. 38.
    Folberg R, Arbieva Z, Moses J, Hayee A, Sandal T, Kadkol S et al (2006) Tumor cell plasticity in uveal melanoma: microenvironment directed dampening of the invasive and metastatic genotype and phenotype accompanies the generation of vasculogenic mimicry patterns. Am J Pathol 169:1376–1389PubMedGoogle Scholar
  39. 39.
    Gildea JJ, Seraj MJ, Oxford G, Harding MA, Hampton GM, Moskaluk CA et al (2002) RhoGDI2 is an invasion and metastasis suppressor gene in human cancer. Cancer Res 62:6418–6423PubMedGoogle Scholar
  40. 40.
    Theodorescu D, Sapinoso LM, Conaway MR, Oxford G, Hampton GM, Frierson HF Jr (2004) Reduced expression of metastasis suppressor RhoGDI2 is associated with decreased survival for patients with bladder cancer. Clin Cancer Res 10:3800–3806PubMedGoogle Scholar
  41. 41.
    Said N, Theodorescu D (2009) Pathways of metastasis suppression in bladder cancer. Cancer Metastasis Rev 28:327–333PubMedGoogle Scholar
  42. 42.
    Titus B, Frierson HF Jr, Conaway M, Ching K, Guise T, Chirgwin J et al (2005) Endothelin axis is a target of the lung metastasis suppressor gene RhoGDI2. Cancer Res 65:7320–7327PubMedGoogle Scholar
  43. 43.
    Wu Y, Siadaty MS, Berens ME, Hampton GM, Theodorescu D (2008) Overlapping gene expression profiles of cell migration and tumor invasion in human bladder cancer identify metallothionein 1E and nicotinamide N-methyltransferase as novel regulators of cell ­migration. Oncogene 27:6679–6689PubMedGoogle Scholar
  44. 44.
    Kedzierski RM, Yanagisawa M (2001) ENDOTHELIN SYSTEM: the double-edged sword in health and disease. Annu Rev Pharmacol Toxicol 41:851–876PubMedGoogle Scholar
  45. 45.
    Herrmann E, Tiemann A, Eltze E, Bolenz C, Bremer C, Persigehl T et al (2009) Endothelin-A-receptor antagonism with atrasentan exhibits limited activity on the KU-19-19 bladder cancer cell line in a mouse model. J Cancer Res Clin Oncol 135:1455–1462PubMedGoogle Scholar
  46. 46.
    Herrmann E, Bogemann M, Bierer S, Eltze E, Toma MI, Kopke T et al (2007) The role of the endothelin axis and microvessel density in bladder cancer – correlation with tumor angiogenesis and clinical prognosis. Oncol Rep 18:133–138PubMedGoogle Scholar
  47. 47.
    Bagnato A, Spinella F, Rosano L (2005) Emerging role of the endothelin axis in ovarian tumor progression. Endocr Relat Cancer 12:761–772PubMedGoogle Scholar
  48. 48.
    Giaid A, Hamid QA, Springall DR, Yanagisawa M, Shinmi O, Sawamura T et al (1990) Detection of endothelin immunoreactivity and mRNA in pulmonary tumours. J Pathol 162:15–22PubMedGoogle Scholar
  49. 49.
    Rosano L, Spinella F, Di Castro V, Nicotra MR, Dedhar S, de Herreros AG et al (2005) Endothelin-1 promotes epithelial-to-mesenchymal transition in human ovarian cancer cells. Cancer Res 65:11649–11657PubMedGoogle Scholar
  50. 50.
    Kandalaft LE, Facciabene A, Buckanovich RJ, Coukos G (2009) Endothelin B receptor, a new target in cancer immune therapy. Clin Cancer Res 15(14):4521–4528PubMedGoogle Scholar
  51. 51.
    Sutcliffe AM, Clarke DL, Bradbury DA, Corbett LM, Patel JA, Knox AJ (2009) Transcriptional regulation of monocyte chemotactic protein-1 release by endothelin-1 in human airway smooth muscle cells involves NF-kappaB and AP-1. Br J Pharmacol 157:436–450PubMedGoogle Scholar
  52. 52.
    Spinella F, Rosano L, Di Castro V, Natali PG, Bagnato A (2004) Endothelin-1-induced prostaglandin E2-EP2, EP4 signaling regulates vascular endothelial growth factor production and ovarian carcinoma cell invasion. J Biol Chem 279:46700–46705PubMedGoogle Scholar
  53. 53.
    Spinella F, Rosano L, Di Castro V, Nicotra MR, Natali PG, Bagnato A (2004) Inhibition of cyclooxygenase-1 and -2 expression by targeting the endothelin a receptor in human ovarian carcinoma cells. Clin Cancer Res 10:4670–4679PubMedGoogle Scholar
  54. 54.
    Wight TN (2002) Versican: a versatile extracellular matrix proteoglycan in cell biology. Curr Opin Cell Biol 14:617–623PubMedGoogle Scholar
  55. 55.
    Domenzain C, Docampo MJ, Serra M, Miquel L, Bassols A (2003) Differential expression of versican isoforms is a component of the human melanoma cell differentiation process. Biochim Biophys Acta 1642:107–114PubMedGoogle Scholar
  56. 56.
    Ricciardelli C, Russell DL, Ween MP, Mayne K, Suwiwat S, Byers S et al (2007) Formation of hyaluronan- and versican-rich pericellular matrix by prostate cancer cells promotes cell motility. J Biol Chem 282:10814–10825PubMedGoogle Scholar
  57. 57.
    Lin HM, Chatterjee A, Lin YH, Anjomshoaa A, Fukuzawa R, McCall JL et al (2007) Genome wide expression profiling identifies genes associated with colorectal liver metastasis. Oncol Rep 17:1541–1549PubMedGoogle Scholar
  58. 58.
    Skandalis SS, Kletsas D, Kyriakopoulou D, Stavropoulos M, Theocharis DA (2006) The greatly increased amounts of accumulated versican and decorin with specific post-translational modifications may be closely associated with the malignant phenotype of pancreatic cancer. Biochim Biophys Acta 1760:1217–1225PubMedGoogle Scholar
  59. 59.
    Hirakawa S, Hong YK, Harvey N, Schacht V, Matsuda K, Libermann T et al (2003) Identification of vascular lineage-specific genes by transcriptional profiling of isolated blood vascular and lymphatic endothelial cells. Am J Pathol 162:575–586PubMedGoogle Scholar
  60. 60.
    Malemud CJ (2006) Matrix metalloproteinases (MMPs) in health and disease: an overview. Front Biosci 11:1696–1701PubMedGoogle Scholar
  61. 61.
    Ricciardelli C, Brooks JH, Suwiwat S, Sakko AJ, Mayne K, Raymond WA et al (2002) Regulation of stromal versican expression by breast cancer cells and importance to relapse-free survival in patients with node-negative primary breast cancer. Clin Cancer Res 8:1054–1060PubMedGoogle Scholar
  62. 62.
    Domenzain-Reyna C, Hernandez D, Miquel-Serra L, Docampo MJ, Badenas C, Fabra A et al (2009) Structure and regulation of the versican promoter: the versican promoter is regulated by AP-1 and TCF transcription factors in invasive human melanoma cells. J Biol Chem 284:12306–12317PubMedGoogle Scholar
  63. 63.
    Brown LF, Guidi AJ, Schnitt SJ, Van De Water L, Iruela-Arispe ML, Yeo TK et al (1999) Vascular stroma formation in carcinoma in situ, invasive carcinoma, and metastatic carcinoma of the breast. Clin Cancer Res 5:1041–1056PubMedGoogle Scholar
  64. 64.
    Kischel P, Waltregny D, Dumont B, Turtoi A, Greffe Y, Kirsch S et al (2010) Versican overexpression in human breast cancer lesions: known and new isoforms for stromal tumor targeting. Int J Cancer 126:640–650PubMedGoogle Scholar
  65. 65.
    Paris S, Sesboue R, Chauzy C, Maingonnat C, Delpech B (2006) Hyaluronectin modulation of lung metastasis in nude mice. Eur J Cancer 42:3253–3259PubMedGoogle Scholar
  66. 66.
    Said N, Theodorescu D (2009) The metastasis suppressor RhoGDI2 suppresses expression of Versican, an invasion associated and macrophage stimulatory molecule. In: Proceedings of the American Association for Cancer Research, 2009 Apr 18–22, Denver, CO. AACR, Philadelphia (PA), 2009. Abstract nr 4810Google Scholar
  67. 67.
    Smith SC, Oxford G, Wu Z, Nitz MD, Conaway M, Frierson HF et al (2006) The metastasis-associated gene CD24 is regulated by Ral GTPase and is a mediator of cell proliferation and survival in human cancer. Cancer Res 66:1917–1922PubMedGoogle Scholar
  68. 68.
    Smith SC, Theodorescu D (2009) The Ral GTPase pathway in metastatic bladder cancer: key mediator and therapeutic target. Urol Oncol 27:42–47PubMedGoogle Scholar
  69. 69.
    Wang H, Owens C, Chandra N, Conaway MR, Brautigan DL, Theodorescu D (2010) Phosphorylation of RalB is important for bladder cancer cell growth and metastasis. Cancer Res 70:8760–8769PubMedGoogle Scholar
  70. 70.
    Wu Z, Owens C, Chandra N, Popovic K, Conaway M, Theodorescu D (2010) RalBP1 is necessary for metastasis of human cancer cell lines. Neoplasia 12(12):969–979PubMedGoogle Scholar
  71. 71.
    Henry DO, Moskalenko SA, Kaur KJ, Fu M, Pestell RG, Camonis JH et al (2000) Ral GTPases contribute to regulation of cyclin D1 through activation of NF-kappaB. Mol Cell Biol 20:8084–8092PubMedGoogle Scholar
  72. 72.
    Chien Y, Kim S, Bumeister R, Loo YM, Kwon SW, Johnson CL et al (2006) RalB GTPase-mediated activation of the IkappaB family kinase TBK1 couples innate immune signaling to tumor cell survival. Cell 127:157–170PubMedGoogle Scholar
  73. 73.
    Pirruccello SJ, LeBien TW (1986) The human B cell-associated antigen CD24 is a single chain sialoglycoprotein. J Immunol 136:3779–3784PubMedGoogle Scholar
  74. 74.
    Kristiansen G, Pilarsky C, Pervan J, Sturzebecher B, Stephan C, Jung K et al (2004) CD24 expression is a significant predictor of PSA relapse and poor prognosis in low grade or organ confined prostate cancer. Prostate 58:183–192PubMedGoogle Scholar
  75. 75.
    Fogel M, Friederichs J, Zeller Y, Husar M, Smirnov A, Roitman L et al (1999) CD24 is a marker for human breast carcinoma. Cancer Lett 143:87–94PubMedGoogle Scholar
  76. 76.
    Aigner S, Ramos CL, Hafezi-Moghadam A, Lawrence MB, Friederichs J, Altevogt P et al (1998) CD24 mediates rolling of breast carcinoma cells on P-selectin. FASEB J 12:1241–1251PubMedGoogle Scholar
  77. 77.
    Lopes EC, Ernst G, Aulicino P, Vanzulli S, Garcia M, Alvarez E et al (2002) Dissimilar invasive and metastatic behavior of vincristine and doxorubicin-resistant cell lines derived from a murine T cell lymphoid leukemia. Clin Exp Metastasis 19:283–290PubMedGoogle Scholar
  78. 78.
    Liu W, Vadgama JV (2000) Identification and characterization of amino acid starvation-induced CD24 gene in MCF-7 human breast cancer cells. Int J Oncol 16:1049–1054PubMedGoogle Scholar
  79. 79.
    Ma YQ, Geng JG (2002) Obligatory requirement of sulfation for P-selectin binding to human salivary gland carcinoma Acc-M cells and breast carcinoma ZR-75-30 cells. J Immunol 168:1690–1696PubMedGoogle Scholar
  80. 80.
    Kristiansen G, Sammar M, Altevogt P (2004) Tumour biological aspects of CD24, a mucin-like adhesion molecule. J Mol Histol 35:255–262PubMedGoogle Scholar
  81. 81.
    Ikenaga N, Ohuchida K, Mizumoto K, Yu J, Kayashima T, Hayashi A et al (2010) Characterization of CD24 expression in intraductal papillary mucinous neoplasms and ductal carcinoma of the pancreas. Hum Pathol 41:1466–1474PubMedGoogle Scholar
  82. 82.
    Lee HJ, Choe G, Jheon S, Sung SW, Lee CT, Chung JH (2010) CD24, a novel cancer biomarker, predicting disease-free survival of non-small cell lung carcinomas: a retrospective study of prognostic factor analysis from the viewpoint of forthcoming (seventh) new TNM classification. J Thorac Oncol 5:649–657PubMedGoogle Scholar
  83. 83.
    Mani SA, Guo W, Liao MJ, Eaton EN, Ayyanan A, Zhou AY et al (2008) The epithelial-mesenchymal transition generates cells with properties of stem cells. Cell 133:704–715PubMedGoogle Scholar
  84. 84.
    Dontu G, Liu S, Wicha MS (2005) Stem cells in mammary development and carcinogenesis: implications for prevention and treatment. Stem Cell Rev 1:207–213PubMedGoogle Scholar
  85. 85.
    Lim SC, Oh SH (2005) The role of CD24 in various human epithelial neoplasias. Pathol Res Pract 201:479–486PubMedGoogle Scholar
  86. 86.
    Liu AY, Roudier MP, True LD (2004) Heterogeneity in primary and metastatic prostate ­cancer as defined by cell surface CD profile. Am J Pathol 165:1543–1556PubMedGoogle Scholar
  87. 87.
    Al-Hajj M, Wicha MS, Benito-Hernandez A, Morrison SJ, Clarke MF (2003) Prospective identification of tumorigenic breast cancer cells. Proc Natl Acad Sci U S A 100:3983–3988PubMedGoogle Scholar
  88. 88.
    Baumann P, Cremers N, Kroese F, Orend G, Chiquet-Ehrismann R, Uede T et al (2005) CD24 expression causes the acquisition of multiple cellular properties associated with tumor growth and metastasis. Cancer Res 65:10783–10793PubMedGoogle Scholar
  89. 89.
    Gassmann P, Kang ML, Mees ST, Haier J (2010) In vivo tumor cell adhesion in the pulmonary microvasculature is exclusively mediated by tumor cell–endothelial cell interaction. BMC Cancer 10:177PubMedGoogle Scholar
  90. 90.
    Kohler S, Ullrich S, Richter U, Schumacher U (2010) E-/P-selectins and colon carcinoma metastasis: first in vivo evidence for their crucial role in a clinically relevant model of spontaneous metastasis formation in the lung. Br J Cancer 102:602–609PubMedGoogle Scholar
  91. 91.
    McDonald B, Spicer J, Giannais B, Fallavollita L, Brodt P, Ferri LE (2009) Systemic inflammation increases cancer cell adhesion to hepatic sinusoids by neutrophil mediated mechanisms. Int J Cancer 125:1298–1305PubMedGoogle Scholar
  92. 92.
    Schmidmaier R, Baumann P (2008) ANTI-ADHESION evolves to a promising therapeutic concept in oncology. Curr Med Chem 15:978–990PubMedGoogle Scholar
  93. 93.
    Laubli H, Stevenson JL, Varki A, Varki NM, Borsig L (2006) L-selectin facilitation of metastasis involves temporal induction of Fut7-dependent ligands at sites of tumor cell arrest. Cancer Res 66:1536–1542PubMedGoogle Scholar
  94. 94.
    Fritzsche J, Hunerbein I, Schumacher G, Alban S, Ludwig R, Gille J et al (2005) In vitro investigation on the selectin binding mechanisms in tumor cell metastasis and their inhibition by heparin. Int J Clin Pharmacol Ther 43:570–572PubMedGoogle Scholar
  95. 95.
    Ludwig RJ, Boehme B, Podda M, Henschler R, Jager E, Tandi C et al (2004) Endothelial P-selectin as a target of heparin action in experimental melanoma lung metastasis. Cancer Res 64:2743–2750PubMedGoogle Scholar
  96. 96.
    Louie E, Nik S, Chen JS, Schmidt M, Song B, Pacson C et al (2010) Identification of a stem-like cell population by exposing metastatic breast cancer cell lines to repetitive cycles of hypoxia and reoxygenation. Breast Cancer Res 12:R94PubMedGoogle Scholar
  97. 97.
    Storci G, Sansone P, Mari S, D’Uva G, Tavolari S, Guarnieri T et al (2010) TNFalpha up-regulates SLUG via the NF-kappaB/HIF1alpha axis, which imparts breast cancer cells with a stem cell-like phenotype. J Cell Physiol 225:682–691PubMedGoogle Scholar
  98. 98.
    Shpitz B, Giladi N, Sagiv E, Lev-Ari S, Liberman E, Kazanov D et al (2006) Celecoxib and curcumin additively inhibit the growth of colorectal cancer in a rat model. Digestion 74:140–144PubMedGoogle Scholar
  99. 99.
    Sagiv E, Arber N (2008) The novel oncogene CD24 and its arising role in the carcinogenesis of the GI tract: from research to therapy. Expert Rev Gastroenterol Hepatol 2:125–133PubMedGoogle Scholar
  100. 100.
    Sagiv E, Starr A, Rozovski U, Khosravi R, Altevogt P, Wang T et al (2008) Targeting CD24 for treatment of colorectal and pancreatic cancer by monoclonal antibodies or small interfering RNA. Cancer Res 68:2803–2812PubMedGoogle Scholar
  101. 101.
    Choi YL, Lee SH, Kwon GY, Park CK, Han JJ, Choi JS et al (2007) Overexpression of CD24: association with invasiveness in urothelial carcinoma of the bladder. Arch Pathol Lab Med 131:275–281PubMedGoogle Scholar
  102. 102.
    Sagiv E, Memeo L, Karin A, Kazanov D, Jacob-Hirsch J, Mansukhani M et al (2006) CD24 is a new oncogene, early at the multistep process of colorectal cancer carcinogenesis. Gastroenterology 131:630–639PubMedGoogle Scholar
  103. 103.
    Sagiv E, Kazanov D, Arber N (2006) CD24 plays an important role in the carcinogenesis process of the pancreas. Biomed Pharmacother 60:280–284PubMedGoogle Scholar
  104. 104.
    Sagiv E, Rozovski U, Kazanov D, Liberman E, Arber N (2007) Gene expression analysis proposes alternative pathways for the mechanism by which celecoxib selectively inhibits the growth of transformed but not normal enterocytes. Clin Cancer Res 13:6807–6815PubMedGoogle Scholar
  105. 105.
    Overdevest JB, Smith SC, Thomas S, Theodorescu D (2009) CD24: target for a novel anti-metastatic therapeutic. In: Proceedings of the American Association for Cancer Research, 2009 Apr 18–22, Denver, CO. AACR, Philadelphia (PA), 2009. Abstract nr 4941Google Scholar

Copyright information

© Springer Science+Business Media B.V. 2011

Authors and Affiliations

  1. 1.Department of UrologyPaul Mellon Urologic Cancer Institute, University of VirginiaCharlottesvilleUSA
  2. 2.Departments of Surgery and PharmacologyUniversity of Colorado Comprehensive Cancer CenterAuroraUSA

Personalised recommendations