Tumor-associated Endo180 requires stromal-derived LOX to promote metastatic prostate cancer cell migration on human ECM surfaces
- 2.1k Downloads
The diverse composition and structure of extracellular matrix (ECM) interfaces encountered by tumor cells at secondary tissue sites can influence metastatic progression. Extensive in vitro and in vivo data has confirmed that metastasizing tumor cells can adopt different migratory modes in response to their microenvironment. Here we present a model that uses human stromal cell-derived matrices to demonstrate that plasticity in tumor cell movement is controlled by the tumor-associated collagen receptor Endo180 (CD280, CLEC13E, KIAA0709, MRC2, TEM9, uPARAP) and the crosslinking of collagen fibers by stromal-derived lysyl oxidase (LOX). Human osteoblast-derived and fibroblast-derived ECM supported a rounded ‘amoeboid-like’ mode of cell migration and enhanced Endo180 expression in three prostate cancer cell lines (PC3, VCaP, DU145). Genetic silencing of Endo180 reverted PC3 cells from their rounded mode of migration towards a bipolar ‘mesenchymal-like’ mode of migration and blocked their translocation on human fibroblast-derived and osteoblast-derived matrices. The concomitant decrease in PC3 cell migration and increase in Endo180 expression induced by stromal LOX inhibition indicates that the Endo180-dependent rounded mode of prostate cancer cell migration requires ECM crosslinking. In conclusion, this study introduces a realistic in vitro model for the study of metastatic prostate cancer cell plasticity and pinpoints the cooperation between tumor-associated Endo180 and the stiff microenvironment imposed by stromal-derived LOX as a potential target for limiting metastatic progression in prostate cancer.
KeywordsBone Cell migration Collagen Fibroblast Osteoblast Prostate cancer
Advanced glycation end-product
Bovine serum albumin
C-type lectin domain
Dedicator of cytokinesis 10
Epithelial to mesenchymal transition
Ephrin type-A receptor 2
Fetal bovine serum
Glyceraldehyde 3-phosphate dehydrogenase
Green fluorescent protein
Horse radish peroxidase
Metastatic bone disease
Myosin light chain-2
Neuronal Wiskott–Aldrich syndrome protein
p21 protein (Cdc42/Rac)-activated kinase 2
Phosphate buffered saline
Pyruvate dehydrogenase kinase, isozyme 1
Ras protein-specific guanine nucleotide-releasing factor 2
Rho associated protein kinase
shEndo180 scrambled control
SRY (sex determining region Y)-box 2
Transforming growth factor-beta-1
Transforming growth factor-beta-1 receptor
Urokinase plasminogen activator receptor associated protein
Metastatic bone disease (MBD) affects approximately 1 million advanced cancer patients per annum in the EU, USA and Japan; and estimates suggest that approximately one fifth of MBD cases result from advanced prostate cancer . MBD is normally accompanied by the presence of additional metastatic lesions in visceral organs. However, in vitro experimental systems used to study putative metastatic targets tend to overlook the precise composition, organization and bioactivity of human bone and visceral tissues. The de novo extracellular matrix (ECM) produced by human trabecular bone osteoblasts is abundant in the minerals, proteins and growth factors found in normal human bone, which provides an accurate biomaterial to study therapeutic targets in the context of MBD [2, 3, 4]. Likewise, human fibroblast-derived ECM has been used to develop more realistic in vitro models of human cancer localized in visceral tissue in which its influence on therapeutic strategies can be considered [5, 6].
Tumor cells can adopt different modes of migration during metastasis. Three modes of tumor cell migration include grouped, bipolar and rounded, which respectively involve: (a) collective ‘epithelioid-like’ cell clusters directed by a leader cell; (b) ‘mesenchymal-like’ translocation of single cells coordinated by forward protrusion and rear retraction of the plasma membrane; and (c) ‘amoeboid-like’ forward translocation of singular spheroidal cells [7, 8]. Tumor cells can switch back-and-forth between different modes of migration in response to external and/or internal cues. This type of morphological plasticity is a feature of the epithelial-to-mesenchymal, mesenchymal-to-amoeboid and collective-to-amoeboid transitions that occur during tumor progression [9, 10]. The ‘amoeboid-like’ cell phenotype predominates at the invasive edge of high-grade tumors  and has been identified as an escape mechanism from some anti-invasive strategies . Tumor cells engaged in this rounded mode of cell migration do not require focal adhesion turnover as they do for bipolar ‘mesenchymal-like’ migration . Instead rounded cell migration is driven by the spatial localization of integrins and cytoskeletal regulators at the posterior plasma membrane [14, 15] and generation of RhoA and Rho kinase associated protein kinase (ROCK)-based actinomyosin contractile signals .
The type I transmembrane collagen receptor Endo180 (CD280, CLEC13E, KIAA0709, MRC2, TEM9, uPARAP) is as a strong prognostic indicator for prostate cancer survival [17, 18]. Within this context Endo180 functions as a modulatory switch for epithelial-to-mesenchymal transition (EMT), and pro-invasive behavior in normal prostate epithelial cells triggered by increased crosslinking and stiffness of the basement membrane following its exposure to advanced glycation end-products (AGEs) [17, 18]. The pro-migratory and pro-invasive role of Endo180 involving the promotion of RhoA-ROCK-based actinomyosin contractility at the cell posterior [17, 18, 19] has been confirmed in a range of tumor and stromal cell types, both in vivo and in vitro, using ectopic over expression, genetic silencing, genetic ablation or targeted blockade of receptor function [17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30]. Given the expression of Endo180 observed in tumor cell foci in metastatic bone lesions , and increased levels of soluble Endo180 in the serum of patients with visceral and bone metastases , we hypothesized that Endo180 can regulate prostate cancer cell plasticity on the bone-like ECM derived from human osteoblasts and visceral tissue-like ECM derived from human fibroblasts.
Lysyl oxidase (LOX) is a copper-dependent amine oxidase that is produced by osteoblasts and fibroblasts to give tissue its structural support and mechanical stiffness by crosslinking the adjacent collagen fibers that they deposit as part of the ECM [32, 33]. LOX plays a fundamental role in metastasis [34, 35, 36, 37], including the formation of the pre-metastatic lesions in bone that are colonized by circulating tumor cells and expand into occult osteolytic metastases . Considering the positive cooperation between tumor-associated Endo180 and AGE-dependent crosslinking and stiffness of basement membrane matrix , we hypothesized that Endo180-dependent prostate cancer cell migration cooperates with LOX-dependent crosslinking of the ECM derived from human osteoblasts and fibroblasts.
Materials and methods
Cells and cell culture
For osteoblast isolation approximately fifty post-operative human trabecular bone chips of 1–2 mm2 were washed thoroughly in PBS to remove hematopoietic cells and incubated for 2 h at 37 °C in 10 ml of 1.2 mg/ml type IV collagenase diluted in DMEM (Invitrogen Ltd. Paisley, UK). Supernatants containing digested cellular components (>107 cells) were harvested and cultured at 37 °C in 5 % CO2 in a 1:1 mix of DMEM and F-12 medium (Invitrogen Ltd.) supplemented with 10 % v/v FBS (First Link UK Ltd., Birmingham, UK), 2 mM l-glutamine, 100 U/ml penicillin, 100 mg/ml streptomycin and 0.25 mg/ml amphotericin B (Invitrogen Ltd.). Primary human osteoblasts and HCA2-hTERT human fibroblasts were maintained in DMEM +10 % v/v FCS, 1 mM penicillin/streptomycin and 2 mM l-glutamine.
PC3, DU145 and VCAP cells were maintained in RPMI medium (Invitrogen Ltd.) +10 % v/v FBS, 1 mM penicillin/streptomycin and 2 mM l-glutamine. Endo180 knockdown and control cells were generated by transfection (Qiagen Superfect) of PC3 cells with the shRNA vector pRNATin-H1.2/Hygro containing coral GFP (Antibodies-Online GmbH, Aachen, Germany) and shEndo180 or non-targeting shEndo180 scrambled control (shSCN) sequence inserts [26, 39] that were evaluated previously to rule out any off-target effects on other pro-migratory and pro-invasive proteins [17, 19]. Transfected cells were selected in fully supplemented DMEM containing hygromycin-B (20 µg/ml) (Santa Cruz Biotechnology Inc., Heidelberg, Germany, UK). PC3 cells were transfected with the pcDNA3-Endo180 or empty pcDNA3 vector using lipofectamine and selected with G418 (0.5 mg/ml), as previously described [26, 39].
Matrix preparation and analysis
Rat type I collagen derived from rat tails was commercially sourced (#354236, BD Biosciences, Oxford, UK) and used at a concentration of 50 µg/ml in 0.02 M glacial acetic acid to coat wells following the manufacturers instructions. For human cell-derived ECM production HCA2 fibroblasts and primary human trabecular bone osteoblasts were seeded at a density of 1.5 × 104 cells per well in 96-well plates (#CLS3595, Corning ® Costar ® ). Confluent cultures of HCA2 fibroblasts were stimulated with 2 mM ascorbate to induce the production of native ECM. Confluent cultures of primary human trabecular bone osteoblasts were stimulated with 100 µM ascorbate, 10 nM dexamethasone and 10 mM β-glycerophosphate to induce mineralized ECM production as previously described [2, 4]. Stimulation media were replaced every 2 days. After 10 days cells were washed with PBS (3 × 5 min) and removed by three successive freeze–thaw cycles (in PBS) and incubation in 1 % w/v sodium deoxycholate for 5 min. Resulting decellularized matrices were washed with PBS (5 × 5 min) before use. Mineralization was determined by von Kossa staining (5 % w/v silver nitrate), as previously described .
BAPN (0.1–1.0 mM) was included in stimulation media of fibroblasts and osteoblasts during the 10-day period of matrix generation to inhibit LOX-dependent type I collagen fiber crosslinking. After 10 days the matrices generated in the presence of BAPN were decellularized. The inhibitory effect of BAPN on collagen crosslinking in fibroblast-derived ECM and osteoblast-derived ECM was ascertained by adapting a previously described method . In brief, decellularized ECM derived from fibroblasts and osteoblasts (untreated or treated with BAPN) were immunostained with rabbit anti-human type I collagen polyclonal antibody (R1038X, Acris Antibodies GmbH, Herford, Germany) and secondary anti-rabbit Alexa Fluor 488-conjugated IgG (Invitrogen Ltd.). Images of the type I collagen fibers present in fibroblast-derived ECM and osteoblast-derived ECM were acquired using Image XpressMICRO (IXM) (Molecular Devices UK Ltd., Wokingham UK) . The curvature ratio (defined as x/y, where x = the total length of each fiber and y = the linear distance between the start and end of each fiber) was calculated using Image J software (arbitrary units) in images of type I collagen fibers. The same images were processed using Metamorph® software (Molecular Devices UK Ltd.) to calculate integrated fluorescent signal intensity per unit area (µm2) using a modification of a protocol used to calculate the integrated fluorescent signal intensity of type I collagen fibers per cell .
Cell morphology and migration assays
Criteria used for scoring different modes of prostate cancer cell migration
Cellular morphology observed during translocation
Criteria used to define the predominant mode of migration
Epithelioid; as part of a moving cell cluster or participation in frequent interactions with adjacent cells
>80 % cells display epithelioid morphology during translocation
Mesenchymal; movement as elongated singular cell with defined leading edge and retraction of trailing uropod
Rounded/bipolar ratio <1.0
Amoeboid; movement as spheroid without a retracting uropod
Rounded/bipolar ratio >1.0
Equal numbers of cells with mesenchymal and amoeboid morphologies
Rounded/bipolar ratio = 1.0
Cells were trypsinized and fixed in 4 % w/v paraformaldehyde (10 min), blocked and permeabilized in immunofluorescence buffer (IFB: 4 % w/v BSA and 1 % v/v FBS) containing 0.2 % w/v saponin. Cells were pelleted and incubated with anti-human Endo180 primary monoclonal antibody (A5/158, E1/183 or 39.10) diluted in IFB (1 h), washed in IFB (3 × 5 min), incubated with Alexa Fluor-555 conjugated secondary antibody diluted in IFB (1 h) and washed IFB (3 × 5 min). Cells were pelleted, resuspended in PBS and assessed by flow cytometry (BD FACS Canto, BD, Oxford, UK). Gating was performed with unstained cells and cells stained with isotype matched IgG.
Protein concentrations in whole cell lysates were determined using a Pierce BCA protein assay kit. Equal amounts of protein were resolved by SDS-PAGE using 7 % w/v polyacrylamide gels and electroblotted onto PVDF membranes, which were incubated at room temperature in blocking buffer (PBS + 5 % w/v BSA) (1 h) then primary antibody (anti-Endo180 A5/158 mAb; anti-GAPDH) diluted in blocking buffer at 4 °C (16 h). After washes in PBS+ 0.1 % v/v Tween®-20 (PBS-T) (5 × 5 min) blots were incubated in HRP-conjugated goat anti-mouse or goat anti-rabbit IgG diluted in blocking buffer (1 h). Blots were washed in PBS-T (5 × 5 min) prior to visualization of immunoreactive bands using chemiluminescence.
Cell adhesion assay
Cell adhesion could not be measured using crystal violet because the fibroblast-derived and osteoblast-derived ECM retained the stain. Instead, cells were seeded at a density of 1.5 × 104 per well of a 96-well plate onto test substrata and incubated for 1 h before washing in PBS and addition of culture medium containing CellTiter-Glo® buffer (Promega, UK) at a ratio of 1:1 and final volume of 200 µl. Plates were mixed vigorously for 2 min to induce cell lysis and the contents of each well transferred to opaque 96-well plates (Corning® Costar®; Z37 185-8; Sigma Aldrich Ltd., Poole, UK). Luminescence was measured on a PHERAstarPlus plate reader (BMG LabTech, Aylesbury, UK).
MTT cell proliferation assay
1.2 × 104 cells per well were seeded onto test substrata in 24-well plates and incubated for 48 h. 5 mg/ml MTT was added and cells incubated for 3.5 h at 37 °C. Media was removed and 300 µl of extraction buffer (0.5 M dimethylformamide; 20 % w/v SDS) added per well followed by incubation for 2 h. 100 µl of buffer was transferred per well to a 96-well plate. Absorbance (570 nm) was measured using a Sunrise plate reader (Labtech International Ltd., Ringmer, UK).
Student’s t test was performed using SPSS 15.0 software; p < 0.05 was considered significant.
Generation of human stromal cell-derived ECM surfaces with LOX-dependent cross links
Rounded metastatic prostate cancer cell migration is favored on human bone matrix
Endo180 is upregulated in metastatic prostate cancer cells in contact with human ECM
Endo180 is required for rounded prostate cancer cell migration on human stromal ECM surfaces
Endo180 cooperates with fibroblast-derived LOX to promote metastatic prostate cancer cell migration
Migratory mode, velocity, adhesion and proliferation rates of human prostate cancer cell lines on human fibroblast and osteoblast-derived matrices
Tissue culture plastic
Commercial type I collagen
The intracellular mechanisms of rounded tumor cell migration delineated so far have been centered upon the suppressor and activator signals that regulate RhoA-ROCK and myosin light chain-2 (MLC2)-dependent actinomyosin-based contractility, cytoskeletal remodeling and dynamic cell adhesion events. For example, it has been demonstrated that rounded cell movement can be reversed by Smurf-1, a E3 ubiquitin ligase that targets RhoA for degradation, and PDK1, which antagonizes the RhoE-dependent activation of ROCK [45, 46]. Rounded cell migration is also driven by aberrant activation of RhoA following loss of p53 and p27, the suppression of Rac1 and SOX2, or the expression of EphA2 [43, 47, 48, 49, 50, 51, 52]. The interaction between RhoC and FMNL2, Cdc42 and its regulators (DOCK10, RasGRF2) and effectors (N-WASP, PAK2), also promotes rounded cell migration [44, 53, 54, 55, 56]; whereas the dephosphorylation of stathmin (a microtubule destabilizing protein), loss of cofilin or depletion of paxillin can block rounded cell migration [57, 58, 59].
In this study we have pinpointed the collagen receptor Endo180 as a novel modulator of rounded tumor cell migration in the context of the bone (osteoblast-derived ECM) and visceral tissue (fibroblast-derived ECM) microenvironments. This novel pro-migratory mechanism is consolidated by the upregulation of Endo180 expression by up to ~20-fold in PC3 cells, ~9-fold in VCaP cells and ~7-fold in DU145 cells on osteoblast-derived ECM compared to control substrata. The possible intracellular cues that can direct this Endo180-associated tumor cell plasticity include Cdc42 and Rac1 and the Rho-ROCK-MLC2 pathway, which are activated by the spatiotemporal localization of the Endo180 receptor to the plasma membrane or constitutively recycling endosomes [18, 19, 26]. Interestingly two key Endo180 interaction partners, CD147 and urokinase-type plasminogen activator receptor (uPAR) [17, 26], have been identified as regulators of rounded cell migration [60, 61]. CD147-annexin II complex acts as a molecular switch that directs rounded-to-bipolar transitions during cell migration. It is feasible that Endo180-CD147 complex  also plays a modulatory role in tumor cell plasticity on human stromal cell-derived ECM. In this respect we hypothesize that Endo180-CD147 complex disruption can promote rounded tumor cell migration and Endo180-CD147 complex formation can uncouple Endo180 and the intracellular machinery that drives rounded tumor cell migration. It is also possible that the integrin-dependent actomyosin contractile signals generated at the pseudo-uropod-like structure at the rear of spheroidal cells during rounded cell migration  involves the spatiotemporal activation of Rho-ROCK-MLC2-based contractile signals by Endo180-containing endosomes. This prediction is supported by the fact that de-adhesion of the uropod at the rear of MG63 osteosarcoma cells requires the Endo180-Rho-ROCK-MLC2 signalling axis . It will also be interesting to consider if the strong Endo180 clusters in PC3-Endo180 cells on osteoblast-derived ECM contribute to their rounded mode of migration.
The requirement of LOX-dependent ECM crosslinking and stiffness for Endo180-dependent tumor cell migration is aligned with the finding that non-enzymatic crosslinking of basement membrane matrix coupled with Endo180-dependent mechanotransduction triggers epithelial cell invasiveness . When considering the design of Endo180 based anti-metastatic therapies it will be important to fully explore the relative contributions of the two functional C-type lectin domains (CTLDs) in the receptor, CTLD2 and CTLD4, to the migratory behavior of metastatic prostate cancer cells in the context of human ECM lattices that have different levels of stiffness. Our findings indicate that where the ECM is more compliant Endo180 and CD147 form a molecular complex that involves CTLD4 and suppresses epithelial cell invasiveness . This suggests that in compliant tissue it would not be desirable to target CTLD4. On the flipside, blockade of CTLD2-dependent mechanotransduction, which can inhibit the epithelial cell invasiveness induced by non-enzymatic crosslinking and increased stiffness of the basement membrane , could be used to prevent rounded tumor cell migration in stiff visceral tissue and bone.
In contrast to the finding that Endo180 is uncoupled from its ability to promote tumor cell migration on compliant (non-crosslinked) fibroblast-derived ECM, no differences were observed in the migration of tumor cells on non-crosslinked versus crosslinked osteoblast-derived ECM. Although our findings suggest that osteoblast-derived LOX does not affect metastatic prostate cancer cell migration, tumor-derived LOX participates in the progression of osteolytic bone metastasis in breast cancer . In the current study we did not consider the cooperative roles of tumor-derived LOX and Endo180 in driving the plasticity of tumor cell movement on human fibroblast-derived and osteoblast-derived ECM surfaces. Consideration of this possibility together with the evaluation of Endo180 and LOX as targets in pre-clinical models of osteolytic bone tumors induced by PC3 and DU145 cells [62, 63, 64] and predominantly osteosclerotic tumors induced by VCaP cells  will be prioritised in our future work. The finding that LOX-dependent crosslinking of human fibroblast-derived ECM is required to promote tumor cell migration, indicates that anti-Endo180 and/or anti-LOX therapy is a feasible therapeutic option for the treatment of visceral tumors surrounded by a stiffened stroma (Fig. 6f).
The findings of this study provide new insight into the consequences of Endo180 upregulation on prostate tumor cells in contact with osteoblasts , positive Endo180 immunostaining of tumor cell foci in metastatic bone lesions  and raised levels of soluble Endo180 in the serum of patients with osseous and/or visceral metastases . The heterotypic interaction of osteoblasts with prostate cancer cells was previously shown to suppress Endo180 expression in the osteoblasts resulting in decreased mineralized collagen production . Here we have demonstrated that osteoblast-derived ECM increases Endo180 expression in tumor cells to drive their transition to a rounded mode of migration. Therapeutic strategies that can suppress Endo180 function in metastatic disease (Fig. 6f), combined with the development of Endo180-targeted diagnostics, could provide the opportunity to make a major advance in the personalized treatment of men with Endo180-positive prostate cancer who are at risk of, or have progressed towards, the development of Endo180-driven bone metastasis [17, 18].
We thank Professor Justin Cobb (Imperial College London) for providing human trabecular bone, Professor David Kipling (Cardiff University) for HCA2-hTERT human fibroblasts and Professor Daniel Aeschlimann for the HCA2 matrix generation protocol. We thank Alexandra Glymond and Imogen S Broadbent (University of Hull) for their assistance with data analysis. The Rosetrees Trust (Grants JS16/M59, M40/M41), The Association of International Cancer Research (now Worldwide Cancer Research) (Grant 08-0803) and The Prostate Cancer Charity (now Prostate Cancer UK) (Grant 110632) funded this work. NS was funded by a British Society of Cell Biology summer studentship.
Compliance with ethical standards
Conflict of interest
The authors declare that they do not have any competing or financial interests.
The use of post-operative human trabecular bone from patients undergoing hip replacement surgery was granted approval by Research Ethics Committee of Imperial College NHS Healthcare Trust (reference number: 10/H0711/19) who specifically approved use in prostate cancer research and all persons gave informed consent prior to inclusion in the study. Therefore this research was performed in accordance with the ethical standards laid down in the 1964 Declaration of Helsinki and its later amendments.
- 2.Reichert JC, Quent VMC, Burke LJ, Stansfield SH, Clements JA, Hutmacher DW (2010) Mineralized human primary osteoblast matrices as a model system to analyse interactions of prostate cancer cells with the bone microenvironment. Biomaterials 31(31):7928–7936. doi: 10.1016/j.biomaterials.2010.06.055 CrossRefPubMedGoogle Scholar
- 3.Hesami P, Holzapfel BM, Taubenberger A, Roudier M, Fazli L, Sieh S, Thibaudeau L, Gregory LS, Hutmacher DW, Clements JA (2014) A humanized tissue-engineered in vivo model to dissect interactions between human prostate cancer cells and human bone. Clin Exp Metastasis. doi: 10.1007/s10585-014-9638-5 PubMedGoogle Scholar
- 4.Caley MP, Kogianni G, Adamarek A, Gronau JH, Rodriguez-Teja M, Fonseca AV, Mauri F, Sandison A, Rhim JS, Palmieri C, Cobb JP, Waxman J, Sturge J (2012) TGFbeta1-Endo180-dependent collagen deposition is dysregulated at the tumour–stromal interface in bone metastasis. J Pathol 226(5):775–783. doi: 10.1002/path.3958 CrossRefPubMedGoogle Scholar
- 11.Sanz-Moreno V, Gaggioli C, Yeo M, Albrengues J, Wallberg F, Viros A, Hooper S, Mitter R, Feral CC, Cook M, Larkin J, Marais R, Meneguzzi G, Sahai E, Marshall CJ (2011) ROCK and JAK1 signaling cooperate to control actomyosin contractility in tumor cells and stroma. Cancer Cell 20(2):229–245. doi: 10.1016/j.ccr.2011.06.018 CrossRefPubMedGoogle Scholar
- 12.Wolf K, Mazo I, Leung H, Engelke K, von Andrian UH, Deryugina EI, Strongin AY, Brocker EB, Friedl P (2003) Compensation mechanism in tumor cell migration: mesenchymal-amoeboid transition after blocking of pericellular proteolysis. J Cell Biol 160(2):267–277. doi: 10.1083/jcb.200209006 PubMedCentralCrossRefPubMedGoogle Scholar
- 13.Carragher NO, Walker SM, Scott Carragher LA, Harris F, Sawyer TK, Brunton VG, Ozanne BW, Frame MC (2006) Calpain 2 and Src dependence distinguishes mesenchymal and amoeboid modes of tumour cell invasion: a link to integrin function. Oncogene 25(42):5726–5740. doi: 10.1038/sj.onc.1209582 CrossRefPubMedGoogle Scholar
- 17.Rodriguez-Teja M, Gronau JH, Minamidate A, Darby S, Gaughan L, Robson C, Mauri F, Waxman J, Sturge J (2015) Survival outcome and EMT suppression mediated by a lectin domain interaction of Endo180 and CD147. Mol Cancer Res 13(3):538–547. doi: 10.1158/1541-7786.MCR-14-0344-T CrossRefPubMedGoogle Scholar
- 18.Rodriguez-Teja M, Gronau JH, Breit C, Zhang YZ, Minamidate A, Caley MP, McCarthy A, Cox TR, Erler JT, Gaughan L, Darby S, Robson C, Mauri F, Waxman J, Sturge J (2015) AGE-modified basement membrane cooperates with Endo180 to promote epithelial cell invasiveness and decrease prostate cancer survival. J Pathol 235(4):581–592. doi: 10.1002/path.4485 CrossRefPubMedGoogle Scholar
- 21.Engelholm LH, List K, Netzel-Arnett S, Cukierman E, Mitola DJ, Aaronson H, Kjoller L, Larsen JK, Yamada KM, Strickland DK, Holmbeck K, Dano K, Birkedal-Hansen H, Behrendt N, Bugge TH (2003) uPARAP/Endo180 is essential for cellular uptake of collagen and promotes fibroblast collagen adhesion. J Cell Biol 160(7):1009–1015. doi: 10.1083/jcb.200211091 PubMedCentralCrossRefPubMedGoogle Scholar
- 23.Ikenaga N, Ohuchida K, Mizumoto K, Akagawa S, Fujiwara K, Eguchi D, Kozono S, Ohtsuka T, Takahata S, Tanaka M (2012) Pancreatic cancer cells enhance the ability of collagen internalization during epithelial-mesenchymal transition. PLoS ONE 7(7):e40434. doi: 10.1371/journal.pone.0040434 PubMedCentralCrossRefPubMedGoogle Scholar
- 25.Mousavi SA, Sato M, Sporstol M, Smedsrod B, Berg T, Kojima N, Senoo H (2005) Uptake of denatured collagen into hepatic stellate cells: evidence for the involvement of urokinase plasminogen activator receptor-associated protein/Endo180. Biochem J 387(1):39–46. doi: 10.1042/BJ20040966 PubMedCentralCrossRefPubMedGoogle Scholar
- 28.Jensen PR, Andersen TL, Pennypacker BL, le Duong T, Engelholm LH, Delaisse JM (2014) A supra-cellular model for coupling of bone resorption to formation during remodeling: lessons from two bone resorption inhibitors affecting bone formation differently. Biochem Biophys Res Commun 443(2):694–699. doi: 10.1016/j.bbrc.2013.12.036 CrossRefPubMedGoogle Scholar
- 29.Curino AC, Engelholm LH, Yamada SS, Holmbeck K, Lund LR, Molinolo AA, Behrendt N, Nielsen BS, Bugge TH (2005) Intracellular collagen degradation mediated by uPARAP/Endo180 is a major pathway of extracellular matrix turnover during malignancy. J Cell Biol 169(6):977–985. doi: 10.1083/jcb.200411153 PubMedCentralCrossRefPubMedGoogle Scholar
- 31.Palmieri C, Caley MP, Purshouse K, Fonseca AV, Rodriguez-Teja M, Kogianni G, Woodley L, Odendaal J, Elliott K, Waxman J, Sturge J (2013) Endo180 modulation by bisphosphonates and diagnostic accuracy in metastatic breast cancer. Br J Cancer 108(1):163–169. doi: 10.1038/bjc.2012.540 PubMedCentralCrossRefPubMedGoogle Scholar
- 37.Levental KR, Yu H, Kass L, Lakins JN, Egeblad M, Erler JT, Fong SF, Csiszar K, Giaccia A, Weninger W, Yamauchi M, Gasser DL, Weaver VM (2009) Matrix crosslinking forces tumor progression by enhancing integrin signaling. Cell 139(5):891–906. doi: 10.1016/j.cell.2009.10.027 PubMedCentralCrossRefPubMedGoogle Scholar
- 38.Cox TR, Rumney RM, Schoof EM, Perryman L, Hoye AM, Agrawal A, Bird D, Latif NA, Forrest H, Evans HR, Huggins ID, Lang G, Linding R, Gartland A, Erler JT (2015) The hypoxic cancer secretome induces pre-metastatic bone lesions through lysyl oxidase. Nature 522(7554):106–110. doi: 10.1038/nature14492 CrossRefPubMedGoogle Scholar
- 41.Korenchuk S, Lehr JE, MClean L, Lee YG, Whitney S, Vessella R, Lin DL, Pienta KJ (2001) VCaP, a cell-based model system of human prostate cancer. Vivo 15(2):163–168Google Scholar
- 47.Oppel F, Muller N, Schackert G, Hendruschk S, Martin D, Geiger KD, Temme A (2011) SOX2-RNAi attenuates S-phase entry and induces RhoA-dependent switch to protease-independent amoeboid migration in human glioma cells. Mol Cancer 10:137. doi: 10.1186/1476-4598-10-137 PubMedCentralCrossRefPubMedGoogle Scholar
- 48.Berton S, Belletti B, Wolf K, Canzonieri V, Lovat F, Vecchione A, Colombatti A, Friedl P, Baldassarre G (2009) The tumor suppressor functions of p27(kip1) include control of the mesenchymal/amoeboid transition. Mol Cell Biol 29(18):5031–5045. doi: 10.1128/MCB.00144-09 PubMedCentralCrossRefPubMedGoogle Scholar
- 52.Taddei ML, Parri M, Angelucci A, Bianchini F, Marconi C, Giannoni E, Raugei G, Bologna M, Calorini L, Chiarugi P (2011) EphA2 induces metastatic growth regulating amoeboid motility and clonogenic potential in prostate carcinoma cells. Mol Cancer Res 9(2):149–160. doi: 10.1158/1541-7786.MCR-10-0298 CrossRefPubMedGoogle Scholar
- 57.Sidani M, Wessels D, Mouneimne G, Ghosh M, Goswami S, Sarmiento C, Wang W, Kuhl S, El-Sibai M, Backer JM, Eddy R, Soll D, Condeelis J (2007) Cofilin determines the migration behavior and turning frequency of metastatic cancer cells. J Cell Biol 179(4):777–791. doi: 10.1083/jcb.200707009 PubMedCentralCrossRefPubMedGoogle Scholar
- 58.Belletti B, Nicoloso MS, Schiappacassi M, Berton S, Lovat F, Wolf K, Canzonieri V, D’Andrea S, Zucchetto A, Friedl P, Colombatti A, Baldassarre G (2008) Stathmin activity influences sarcoma cell shape, motility, and metastatic potential. Mol Biol Cell 19(5):2003–2013. doi: 10.1091/mbc.E07-09-0894 PubMedCentralCrossRefPubMedGoogle Scholar
- 60.Margheri F, Luciani C, Taddei ML, Giannoni E, Laurenzana A, Biagioni A, Chilla A, Chiarugi P, Fibbi G, Del Rosso M (2014) The receptor for urokinase-plasminogen activator (uPAR) controls plasticity of cancer cell movement in mesenchymal and amoeboid migration style. Oncotarget 5(6):1538–1553PubMedCentralCrossRefPubMedGoogle Scholar
- 61.Zhao P, Zhang W, Wang SJ, Yu XL, Tang J, Huang W, Li Y, Cui HY, Guo YS, Tavernier J, Zhang SH, Jiang JL, Chen ZN (2011) HAb18G/CD147 promotes cell motility by regulating annexin II-activated RhoA and Rac1 signaling pathways in hepatocellular carcinoma cells. Hepatology 54(6):2012–2024. doi: 10.1002/hep.24592 CrossRefPubMedGoogle Scholar
- 63.Akech J, Wixted JJ, Bedard K, van der Deen M, Hussain S, Guise TA, van Wijnen AJ, Stein JL, Languino LR, Altieri DC, Pratap J, Keller E, Stein GS, Lian JB (2009) Runx2 association with progression of prostate cancer in patients: mechanisms mediating bone osteolysis and osteoblastic metastatic lesions. Oncogene 29(6):811–821. doi: 10.1038/onc.2009.389 PubMedCentralCrossRefPubMedGoogle Scholar
- 65.Graham TJ, Box G, Tunariu N, Crespo M, Spinks TJ, Miranda S, Attard G, de Bono J, Eccles SA, Davies FE, Robinson SP (2014) Preclinical evaluation of imaging biomarkers for prostate cancer bone metastasis and response to cabozantinib. J Nat Cancer Inst 106(4):dju033. doi: 10.1093/jnci/dju033 CrossRefPubMedGoogle Scholar
Open AccessThis article is distributed under the terms of the Creative Commons Attribution 4.0 International License (http://creativecommons.org/licenses/by/4.0/), which permits unrestricted use, distribution, and reproduction in any medium, provided you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made.