Occupy tissue: The movement in cancer metastasis.

2012·0.14 MB·English

by  Bradbury, Peta

CellAdhesion&Migration6:5,424–432;September/October2012;G2012LandesBioscience
Occupy tissue
The movement in cancer metastasis
Peta Bradbury,1,2 Ben Fabry3 and Geraldine M. O’Neill1,2,*
1Children’sCancerResearchUnit;KidsResearchInstitute;TheChildren’sHospitalatWestmead;Sydney,NSWAustralia;2DisciplineofPaediatricsandChildHealth;
TheUniversityofSydney;Sydney,NSWAustralia;3DepartmentofPhysics;UniversityofErlangen-Nuremberg;Erlangen,Germany
Keywords: migration, invasion, metastasis, adhesion, 3D, 2D
Abbreviations: 2D, 2-dimensional; 3D, 3-dimensional; ECM, extracellular matrix; CDM, cell-derived matrix
Interstitial Dissemination
Thecriticalroleofmigrationandinvasionincancermetastasis
warrantsnewtherapeuticapproachestargetingthemachinery
regulating cell migration and invasion. While 2-dimensional The ability of cancer cells to invade tissue surrounding the
(2D) models have helped identify a range of adhesion primary tumor site, leading to the development of metastatic
molecules, cytoskeletal components and regulators that are tumors, is one of the hallmarks of cancer.1 Disseminating tumor
potentiallyimportantforcellmigration,theuseofmodelsthat cellsencounteravarietyoftissuearchitecturesthroughwhichthey
bettermimicthe3-dimensional(3D)environmenthasyielded must transmigrate. While the majority of tumor cells are
new insights into the physiology of cell movement. For confronted by collagen-rich connective tissue found around most
example, studying cells in 3D models has revealed that
organs in the body, invasion in the brain represents a specialized
invading cancer cells may switch between heterogeneous
form of transmigration due to the unique makeup of the brain
invasionmodesandthusevadepharmacologicalinhibitionof
interstitium. For more detail we direct readers to an excellent,
invasion.Herewesummarizepublisheddatainwhichtherole
recent review that provides a comprehensive description of tissue
of cell adhesion molecules in 2D vs. 3D migration have been
structures through which cancer cells transmigrate in vivo.2 The
directly compared and discuss mechanisms that regulate
migration speed and persistence in 2D and 3D. Finally we structureandcompositionoftheinterstitialtissuesdeterminesthe
discusslimitsof3Dculturemodelstorecapitulatetheinvivo mechanisms that must be employed by the cancer cells for
situation. successful navigation. Gritsenko and colleagues2 suggest three
generic mechanisms employed by invasive cancer cells: contact
guidance along stiff matrix elements through integrin mediated
mechanisms, directed migration toward chemokines and growth
factors and physical pushing through small spaces.
Introduction
Cell Migration In Vivo
Nearly all adherent cells are able to crawl and migrate under
standard 2D cell culture conditions. Time-lapse recordings of The process of cell migration and invasion is, of course, not
migrating cells never fail to excite a sense of awe about the restricted to metastatic cancer. Cell migration and invasion
complexityofthemigrationprocess,aslamellipodiaandfilopodia occurs during embryonic development and continues through-
form, extend and make contact with the matrix, as cells change out adulthood as it is critical for the correct execution of a
their shape, lunge forward, contract and detach. Cell migration variety of biological programs. During embryonic development
depends on a variety of temporally and spatially orchestrated there are precise migration and invasion events that are
parameters, with several hundreds of proteins involved. Although necessary for the final correct organization of the adult tissue.
changesincellmigrationafterknockoutormutationofaprotein A particularly well-studied example of this is during the
may not be the ultimate proof of a causal relationship; watching development of the mammalian brain, where cells undergo
cells migrate and measuring parameters of cell speed and precisely timed migration events that are essential for the final
directional persistence have proven invaluable in elucidating the tissue organization and wiring of the fully developed brain (for
functional components regulating intrinsic cell migration in 2D. review, see ref. 3). The migration speed and persistence of
A number of studies have now begun to examine these same distinct cell populations appear to be important parameters for
parameters using 3D culture systems. determining their final destination in the maturing brain tissue.
Moreover, cell populations change speed as they migrate
throughout different zones of the brain.4 Mutations that alter
*Correspondenceto:GeraldineM.O’Neill;
speed and directional persistence of migrating neuronal cell
Email:[email protected]
populations can therefore perturb the organization of the brain
Submitted:06/08/12;Revised:07/19/12;Accepted:07/20/12
http://dx.doi.org/10.4161/cam.21559 tissue into discrete layers.5,6
424 CellAdhesion&Migration Volume6Issue5
REVIEW
The consequences for changes in speed and persistence of Eventhen,comparativeanalysisofcellmigrationandmorphology
invasive cancer cells are currently less well known. Approaches has revealed striking inconsistencies between cells grown on 2D
that abrogate cell movement in environments that mimic the surfaces vs. those seeded within 3D matrices.13,18 Therefore, in
tissueorganizationoftheinvivotumoraregenerallyconsideredto attempting to recreate a cell’s in vivo environment, it is not
bestrongcandidatesfornoveltherapiestotreatmetastaticcancer. sufficient to only consider which ECM group the cell type in
Similarly, approaches that donotcompletely abrogate movement questionismostlikelytointeractwith;itisequallyimportantthat
but instead slow down the rate of cell migration are expected to the 3D structure and mechanical properties of the matrix closely
restrict the dissemination of cells from the primary tumor and mirror that of the in vivo environment. Three of the most
increase the window of opportunity for localized and targeted commonly used in vitro 3D matrix models—fibrous gels,
delivery of therapies. In the particular case of highly invasive basement membrane extracts (or matrigel) and cell-derived
primary brain tumors such as the deadly grade IV gliomas (also matrices—are discussed in the following sections.
knownasglioblastoma),thereisextensivecollateraldamagetothe Fibrous gels. Fibrous gels, namely reconstituted collagen and
brain tissue as a result of the invading tumor cells, leading to fibrin gels, aim to replicate the major architectural and structural
significantlossofqualityoflife.7Thus,evenapartialreductionof components of the extracellular matrix. Collagen and fibrin
tumorinvasionmayamelioratesomeofthecollateraldamagethat emerged as ideal candidates for single-protein 3D matrix models
occurs in the brains of these patients. Moreover, targeting the as both are major structural components of the extracellular
cell’s migration machinery may in turn sensitize the cells to matrix. It is worth noting, however, that interstitial tissue is
different therapies. One example of such sensitization is tumor composedofheterogenousmixturesofECMcomponents,which
cells lacking expression of the invasion/metastasis promoter these gels do not recapitulate.19 Nevertheless, due to the
NEDD9. These cells appear to concomitantly become sensitized abundance of collagen and fibrin in vivo, reconstituted fibrous
to the effects of Src kinase inhibition.8 Similarly, decreasing the gels are physiologically compatible with numerous cells types.
persistence of the cancer cell movement will necessarily decrease Fibrin gels are particularly suitable to study migratory events
the numbers of successful escapers from the primary tumor site. involved in wound healing,20,21 while collagen gels are more
But to confirm these speculations and expectations, any anti- suitable to study interstitial cell migration and metastatic
invasivetherapymustfirstdemonstrateitseffectivenessinmodels invasion.10,22,23
that mimic the natural in vivo 3D organization of the relevant Notallfibrousgelsarecreatedalike,however.Seeminglyminor
tissue surrounding a particular tumor. protocol variations can have major effects on the gel properties.
For instance, an increase in fiber concentration enhances fiber
3D Culture Models density,24,25 resulting in an overall increase in the mechanical
strength of the gel.26 Additionally, the effective pore size of the
The interactions between cells and the external environment, fibrous meshwork decreases at higher fiber concentrations,27 thus
either with the ECM or with neighboring cells, are in a literal increasing steric hindrance for migrating cells.28,29 The gelation
sense vitally important. Once thought to solely provide physical temperature will also affect the mechanical and structural details
and structural support, the ECM is now known to regulate and ofthegel;forinstance,theporesizeandfibrildiameterdecreases
influence survival,9 proliferation,9 migration and adhesion,10,11 and the number of fibrils formed increases at higher polymeriza-
morphology,12 internal cellular structures13 and signaling path- tion temperatures.27,30
ways.11 During tumorigenesis, cellular interaction with the ECM The production details of harvesting, dissolving and purifying
canbecomederegulatednotonlyasaresultofalteredintegrinand collagenorfibrinfromdifferentanimaltissuesarealsoasourceof
focaladhesionproteinexpression,butalsobecausetumorcellscan substantial variability. This is especially true in the case of
rearrange the local native tissue architecture, alter its chemical collagen, wherefibersharvestedforexamplefrombovine Achilles
composition,structureandstiffness,whichinturnoftenpromotes tendon demonstrate a higher denaturation temperature and a
cellmigrationandinvasion.14,15Forexample,inbreastcarcinomas thicker fiber diameter when compared with collagen harvested
the formation of the distinctive palpable “lump,” known as from rat tail tendon.27,31,32 The method for dissolving the
desmoplasia, is attributed to tumor cell secretion of platelet collagen, whether with acid or by proteolytic digestion (i.e.,
derived growth factor (PDGF). This induces the deposition of pepsin or trypsin) similarly influences the structure and
large quantities of collagen and collagen cross-linkers by polymerization kinetics of the reconstituted gels. Acid-dissolved
surrounding fibroblasts, thus increasing local tissue stiffness and collagen will begin the fibrillogenesis process more rapidly,31,33,34
rigidity.16,17 showing characteristic fiber lengths and diameters that closely
Invivo,mostinvadingcancer cells encountertwomajorECM resemble those observed in vivo.35 In comparison, collagen
groups.Thefirstisthebasementmembranethatdirectlyinteracts extracted by proteolytic enzyme digestion shows significantly
with both the epithelium and endothelium layers. The second is smaller fiber diameters and reduced fibril formation, thus
theinterstitial matrix.Itharbors numerouscollagenisoforms and reducing the gels’ strength and rigidity.30,31,36
fibronectinandisthoughttocontributetotheoverallmechanical Basement membrane extracts. Matrigel is a reconstituted
strength of the tissue. In vitro, the extracellular matrix is most basement membrane matrix product extracted from Engelbreth-
commonlyplanar,tissueculture-treatedplasticorsometimesglass Holm-Swarm (EHS) mouse sarcoma cells. When polymerized,
coatedwithcollagen,fibronectin orotherECM-derived proteins. matrigel mimics the microenvironment of in vivo extracellular
www.landesbioscience.com CellAdhesion&Migration 425
basement membrane.37-39 Composed chiefly of laminin, collagen but decreasing speeds in 2D.57 Without exception, the knock-
IV isoforms and heparan sulfate proteoglycans, basement down or loss of cytoplasmic members of focal adhesion sites
membranes provide a unique signaling platform for cells that (p130Cas, NEDD9, vinculin, talin, FAK, VASP, paxillin and
adhere to it.40,41 Matrigel 3D substrates can restore the normal Hic-5) inhibited cell migration speed in 3D cultures, yet again
morphological characteristics and specific cell functions that varied results are seen in 2D cultures. Depletion of these
would otherwise be lost under standard cell culture conditions, molecules variously increased 2D speed (p130Cas, NEDD9 and
especially in epithelial-,42,43 endothelial-44 and Schwan-derived vinculin knockout fibroblasts), decreased 2D speed (talin and
cells.45 But this is not generally true for all cells; for instance FAK) or did not alter speed relative to controls (vinculin shRNA
fibroblasts that are not normally in contact with a basement in HT1080 fibroblasts and VASP). With the exception of EGF
membrane in vivo adopt a noticeably uncharacteristic rounded stimulation and high a5β1 expression, all treatments resulted in
morphology and exhibit a non-migratory phenotype when reduced intrinsic persistence of migration in 3D. As was seen for
cultured in matrigel.12,39 the speed data, there is no clear correlation between effects on
Cell-derivedmatrix.Whenculturedathighdensity,fibroblasts migration persistence in 2D and effects seen in 3D.
“bioengineer” their own extracellular matrix.46 This cell-derived
matrix(CDM)iscomposed of fibronectin,47 collagentypes Iand Forces in Cell Migration
II,48 hyaluronic acid49 and heparin sulfate proteoglycans.49,50
CDMs mirror the flexibility and malleability of an in vivo The disparate results for 2D and 3D speed and persistence in
ECM, as cells seeded onto a CDM can reorganize and modulate exactly the same cell background highlight critical differences
the matrix, recapitulating the migration and invasion events of between these different model systems. Cell migration in 2D
both physiological and metastatic processes observed in differsinonefundamentalaspectfrom3Dmigrationinthatsteric
vivo.11,12,51 However, as with all protocols, there are limitations hindrance is absent. In 2D, the cell needs to overcome only the
to this method also. Somewhat problematic is the poorly defined frictional (drag) forces from the surrounding liquid and the 2D
andhighlyvariablecompositionoffibroblasticCDMs.Moreover, surface.At aspeedof only afew microns per minute,liquid drag
althoughCDMsdisplaya3Dmatrixorganization,thesematrices can be neglected, as the force of a single myosin motor would be
are typically thin and therefore subject to the rigidity of the sufficienttopropelthecellforward.Thus,thebetteracelladheres
underlying2Dsurface.Regardless,theuseofCDMhaspermitted to its substrate, the greater the contractile force needed to
insights into the signaling pathways that govern proliferation, overcome the adhesive friction from cell-matrix adhesions; as a
morphology, cell-matrix attachments and individual cell migra- general rule, a cell with poorer adhesion can migrate faster on a
tion, all of which impact and promote tumorigenesis.11,12,51 2D surface. Poorer adhesion can either be a consequence of a
lowermatrixproteindensityonthesurface,58oraconsequenceof
Cell Adhesion a reduced expression level of adhesion proteins on the cell (for
examples, see Table 1). Of course, a minimum amount of
The role of focal adhesions has been thoroughly studied in 2D adhesion is essential for cell movement as otherwise the cell
culturesandhasprovidedimportantinsightintothemechanisms cannot polarize or activate its contractile machinery.58 Thus, in a
ofcellmovementin2D.However,asinvestigatorshaveturnedto 2D context, changes in focal adhesion dynamics59,60 dominantly
the analysis of focal adhesions in 3D, there has been conflicting effect cell migration speed and likely explain the disparate effects
reports regarding their presence and detection in 3D.12,52,53 We on 2D cell migration seen following depletion of focal adhesion-
direct readers to the excellent review by Harunaga54 for a associated molecules (Table 1).
comprehensiveanalysisoftheissueofcell-ECMadhesions in3D In 3D, in addition to frictional forces, the cell also has to
culture models. The upshot of which is that focal adhesions do overcomeforcesthatarisefromthesterichindranceofthematrix
exist in 3D, but the composition, mechanical properties and network, provided that the pores and crevices through which the
structure ofthe3Dmatrixprofoundlyaffects theappearance and cellmigratesaresmallerthanthecellitself.Here,thecellhastwo
distribution of focal adhesions.12 options. It can either deform itself until it can fit through the
InTable1 wesummarizedatafromstudiesinwhichmigration pores; or it can deform the network until the pores are large
parameters of speed and persistence have been directly compared enough for passage. Switching between cell body deforming vs.
between 2D and 3D culture conditions in the same cell matrix deforming migration strategies is evidenced by changes in
background. Treatments that are expected to promote invasion cell morphology from rounded cell shapes to elongated cell
and metastasis in vivo such as stimulation with EGF, complete shapes.61Deforming thecell bodysufficientlytosqueezethrough
oncogenic transformation following combined expression of smallporesrequiresforcesforovercomingtheelasticandfrictional
ErbB2 and 14-3-3f and high a5β1 expression all promoted forces of the cytoskeleton. The cell can decrease cytoskeletal
faster migration speed in 3D cultures, yet had variable effects in elasticity (stiffness) and friction by depolymerizing cytoskeletal
2D (Table 1). For example, breast epithelial cells induced to filaments, but although this would reduce the forces necessary to
undergo oncogenic transformation displayed reduced migration deform the cell until it fits through a pore, it also reduces the
speed in 2D,55 while EGF treatment of glioblastoma cells force-generatingcapacityoftheacto-myosincontractileapparatus.
increased 2D speed.56 Similarly, cells from a metastatic Analogously, deforming the matrix network requires forces for
progression series showed increasing speeds in 3D collagen gels, overcoming the elastic and frictional forces of the matrix. By
426 CellAdhesion&Migration Volume6Issue5
Table1.Studiesreportingcellspeedandpersistencein3Dculturescomparedwith2Dmigrationparametersinthesamecellbackground
Adhesion Cellline 2D 3D
molecule
Speed Persistence Speed Persistence
p130CasshRNA52 HT1080 ↑ ↑ 3Dcollagen ↓
fibrosarcoma ↓
NEDD9knockout60,† Fibroblast ↑ ND 3Dcollagen ↓
↓
Vinculinknockout87 Fibroblast ↑ ↑ 3Dcollagen ND
↓
VinculinshRNA52 HT1080 — — 3Dcollagen ↓
fibrosarcoma Nochange
b3integrin Fibroblast ↑ ↓ CDM# ↓
knockout59,* ↑
EGFstimulation56 U87MG ↑ ↓ 3Dcollagen|| 3Dcollagen
glioblastoma ↑ ↑
b1integrin DU-145 Biphasic ND Lowmatrigel%: ND
inhibition52 prostatecancer ↑
Highmatrigel%:
↓
TalinshRNA52 HT1080 ↓ ↑ 3Dcollagen ↓
fibrosarcoma ↓
FAKshRNA52 HT1080 ↓ ↑ 3Dcollagen ↓
fibrosarcoma ↓
VASPshRNA52 HT1080 — — 3Dcollagen ↓
fibrosarcoma ↓
Transformation MCF10anon- (1)↑ (1)↓ “Compliant” “Compliant”3Dcollagen4:
progression transformed (2)↓ (2)↓ 3Dcollagen4: (1)—
series:55 breastepithelia (3)↓ (3)↓ (1)— (2)—
(1)+ErbB2 (2)— (3)↓
(2)+14-3-3f (3)↑ “Stiff”
(3)+ErbB2+14-3-3f “Stiff” 3Dcollagen1:
3Dcollagen1: (1)↓
(1)↓ (2)↓
(2)↓ (3)↓
(3)↑
Metastasisprogressionseries:57 NmuMgmouse ↓ ↓ Collagengel: Collagengel:
(1)67NR—tumorigenic breastepithelial (relative (relative ↑ 66c14
(2)168FARN—micrometastases line to to (relativeto ↓
(3)4T07—secondarymicrometastases 67NR) NmuMG) 67NR)
(4)66c14—secondaryTumors
PaxillinsiRNA94," MDA-MB-231 ND ND CDM# ↓
breastcancer ↓
Hic-5siRNA94 MDA-MB-231 ND ND CDM# ↓
breastcancer ↓
Higha5b162 MDA-MB-231 ND ND 3Dcollagen ↑
breastcancer ↑
*Reducedfocaladhesionlengthandincreasedratesoffocaladhesionturnover;†increasedratesoffocaladhesiondisassembly;4elasticmodulus=103Pa;
1elasticmodulus=391Pa;||effectswerecomparedacrossarangeofcollagenconcentrations(2,3and4mg/mL);"measureddynamicsofGFP-taggedtalin
adhesionsinCDMandshowedmultipleroles—paxillindepletioncausedincreasednumbersofhighlydynamic,peripheral,short-livedadhesions,increased
stability of more centrally located adhesions accompanied by reduced rates of adhesion assembly and disassembly. #cell-derived matrix; ND, not
determined;—,nochange.
secreting matrix-digesting enzymes, the cell can soften the cell migration in 3D give clues as to whether cells are using the
network structure or increase its porosity, thereby reducing the rounded (cell body deforming) vs. elongated (matrix deforming)
necessary forces for deforming the matrix network until the cell cellmigrationstrategies?Initialdescriptionssuggestedthatthereis
canfitthrough.Butatthesametimethisalsolimitstheabilityof nolossofcellspeedbetweenthetwomodes.10However,absolute
the cell to adhere to the matrix, which is a prerequisite for cell speed depends on the make-up of the surrounding matrix.
contractileforcegeneration.Therefore,doesthespeedofintrinsic While partial transformation of breast epithelial cells (Erb2 or
www.landesbioscience.com CellAdhesion&Migration 427
14-3-3f overexpression) did not affect cell speed in “compliant” initialtumor formation.A limitationisthatthis doesnotaddress
collagen gels (103 Pa elastic modulus), cells had reduced initial migration and invasion away from primary tumor sites. A
migrationspeedin“stiff”collagengels(391Paelasticmodulus).55 further important caveat to all of these models is that none
Similarly, increasing steric hindrance at high collagen concentra- measure migration and invasion alone. The ability of the tumor
tions can inhibit cell invasion.62 Thus the concentration of the cellstodisseminateandformsecondarytumorsisamultifactorial
matrix, extent of cross-linking and matrix pore size all critically process, which additionally requires survival in the circulation,
determine cell speed and moreover, whether a matrix deforming adhesion to vessel endothelial layers and transmigration through
or cell body deforming mode of invasion is possible. Given the theendothelium.Belowwecomparedatafor3Dmigrationspeed
ultimate goal of determining the molecular mechanisms that and persistence with in vivo metastasis results in response to
promotecancercellinvasionandmetastasis,itwillbeincreasingly altered adhesion molecule expression and activities.
important toadopt arange of3Dculture modelsthatcan mimic FAK,p130Cas,talinandvinculin.Todate,alargenumberof
the various different extracellular environments encountered by studies have focused on the role of FAK for in vivo metastasis.
invading cancer cells in vivo. Two examples from recent studies FAK has long been known to be upregulated in awide variety of
withopposingfindingshighlighttheproblemininterpretingdata invasive cancer types (reviewed in ref. 86). Cancer cells in which
that are measured under only a single condition: Several studies FAK is either homozygously deleted or downregulated via
have reported that invasive, metastatic cancer cells are softer than expression of a dominant negative form of FAK universally
theirnonmetastaticcounterpartsderivedfromthesametissue63,64 display reduced metastatic tumor formation (Table 2), matching
when measured under non-adherent conditions. By contrast, in the findings that FAK depletion reduces 3D cell migration speed
adherent cancer cells, the opposite behavior is seen, with more (Table1).FAKdepletioninhibitedmetastatictumorformationin
invasive cells tending to be more contractile and consequently all three categories of in vivo invasion models (see above).
stiffer.65,66 Congruent results were also seen between 3D culture migration
speedandinvivometastasesforp130Casandtalin(Tables1 and
Predictive Power of 3D Invasion Models 2).However,thedataarenotsoclearforvinculin.Thedepletion
for Cancer Metastasis orabsenceofvinculinhadnoeffect52orledtoreducedmigration
speed87 in 3D models. In contrast, a high level of exogenous
The goal of the 3D cell migration models is to better mimic the vinculin expression reduced lung tumor formation in vivo.80
physiological environment. An important question in cancer NEDD9. NEDD9 overexpression promotes invasion and
research therefore is how successfully data from the 3D models metastasis in a range of cancer cell types.88 In apparent
predicts in vivo metastatic behavior. To address this question we concordance with this, NEDD9 depletion reduces fibroblast
have searched the literature for examples where adhesion migration in 3D collagen gels (Table 1). However, while
moleculesthathavebeendirectlycomparedin2Dvs.3Dmodels NEDD9−/− tumor cells in a model of oncogenic transformation
(Table 1) have been analyzed using in vivo models of metastasis of mammary epithelia had a tendency to form fewer metastatic
andinvasion(Table2).Thereisanextensiveliteratureonmouse tumors,67 tail vein injection of these isolated cells revealed
models of cancer cell invasion, but for our purposes we have aggressive lung tumor formation.8 Potentially, this may represent
limited our analysis to studies where adhesion molecules from a switch to the amoeboid/cell deforming invasion mode that
Table1 havebeenmanipulatedbyoverexpressionorknockdown. occurs following NEDD9 depletion,22 which may promote lung
It is noticeable that surprisingly few studies have been performed colonization. By contrast, the dense, rigid matrix that is reported
to directly test the effects of these molecules on in vivo tumor to characteristically surround breast tumors,89 which is presum-
invasion and metastasis—the bulk of studies have focused on ably present in the transformed breast epithelia model, may
tumorinitiation.OursearchyieldedexamplesforFAK,p130Cas, obviate the rounded/cell deforming mode of invasion.
NEDD9,vinculin,talin,β1integrinanda5β1andavβ3integrin Alternatively, increased metastatic lesions following tail vein
receptors (Table 2). injection may reflect changes in survival in circulation, adhesion
The in vivo models used in the summarized studies can be and transmigration of the endothelial cell layer of the vessel.
divided into three categories: (1) tissue-specific neo-oncogenesis β1 integrin. The effects of β1 integrin depletion on migration
that recapitulates carcinoma progression seen in human dis- in matrigel are dependent on the matrigel concentration; at high
ease67-72 and orthotopic injection: both have the advantage of concentrations when the gel is less deformable, the loss of β1
mimicking the correct tissue context of the human disease,73-77 impaired the ability of the cells to invade.90 These results are in
(2) sub-cutaneous injection at non-orthotopic sites78-80 and agreement with in vivo data where β1 depletion also caused
(3) intravenous injection.8,81-85 In the case of the first two reduced metastatic lesions (Table 2). In one of the studies,
categories successful metastasis requires the tumor cells to invade increased numbers of tumor emboli in the lymphatic vasculature
locally, intravasate into blood or lymph vessels, extravasate at were noted.68 This suggests that the cells in this model were
secondary sites and form new tumors. Conversely, intravenous competent to locally invade and intravasate, but were not able to
injection does not depend on the formation of a primary tumor, execute the final stages required for secondary tumor formation.
thusallowingseparationoftheeffectsofprimarytumorformation a5β1and avβ3integrinreceptors.Thefindingthathighlevel
from dissemination into secondary tissues. This approach is a5β1isassociatedwithenhancedmigrationspeedandpersistence
frequentlyusedwhenthemoleculeorpathwayofinterestinhibits of breast cancer cells in 3D collagen gels62 appears to agree with
428 CellAdhesion&Migration Volume6Issue5
Table2.Summaryofinvivometastasisdata,targetingtheadhesionmoleculesdescribedinTable1
Adhesion Invivomodel Metastasisphenotype
molecule
FAK Intravenousinjection Failedtoformlungtumors;cellsretainedinthelungcapillarybedwere
ofFAK−/−tumorcells82 roundedandlackedmembraneextensionsintothevessel
ActivatedErbB2mammarytumormodelwith Metastaticlungtumorsallnegativeforcellswithhomozygousdeletionof
epithelial-specificFAKdeletion69 FAK
Mammaryepithelium-specificdeletionofFAKin Metastaticlungtumorsallnegativeforcellswithhomozygousdeletionof
MMTV-PyVmTMouseTumorModel70 FAK
Mammaryepithelium-specificdeletionofFAKin Reducedlungtumormetastases
MMTV-PyVmTMouseTumorModel71
Orthotopicinjectionofpancreaticcancercellstreatedwith Preventedformationoflivermetastases
FAKsiRNA73
p130Cas Mammarytumormodel—injectionofcellsexpressing Inhibitslungcolonization
induciblep130CasshRNA77
Athymicnudemiceinjectedsub-cutaneouslywith Exogenousp130Casexpressionincreasedformationofmetastaticlung
p130Cas−/−fibroblaststransformedwithoncogenic tumorsaftersurgicalremovalofprimarytumors;authorscommentthat“the
Srcandexpressingp130Cas78 capacityofthecellstoinvadethroughmatrigelwasstronglycorrelatedwith
theircapacitytoinvadeandmetastasizeinvivo”
Vinculin Exogenousvinculinexpressioninhighlymetastaticrat Highestlevelsofvinculinexpressionsuppressedformationoflung
adenocarcinomainjectedintofootpad80 metastases,lowtomoderateexpressorsformedtumorsinlymphnodes
closetoinjectionsitebutfailedtoformlungmetastases
NEDD9 Mammaryepithelium-specificdeletionofNEDD9in Trendtofewerlungmetastases
MMTV-PyVmTMouseTumorModel67
TailveininjectionofNEDD9-nullprimarytumors8 Tumorsformedofnull-celllinesexhibitedincreasedaggressiveness,withall
injectedmicegeneratingsecondarytumors
Talin Tailveininjectionsofprostatecancercelllinestreated Reducednumbersofmetastaticlunglesions
withtalinshRNA84
avb3 Orthotopicinjectionintomammaryfatpadwithmammary Droveuniqueformationofbonemetastases;authorsshowincreased
carcinomacelllineexpressingexogenousb376 haptotacticandchemotacticresponsetobone-matrixproteinsand
solublefactors
MDA-MB-435breastcancercellsexpressingconstitutively Enhancedlungcolonization
activeavb3injectedintomousetailvein81
Intravenousinjectionofmetastaticavb3negative Re-expressionofb3inmetastatic,b3negativelinesreducedlung
melanomacellsexpressingexogenousb391 colonization
b1integrin Conditionaldeletionofb1integrinfrommammaryepithelia, Significantlyreducedformationoflungmetastases
crossedwithMMTV/activatederbB272
Orthotopicinjectionofpancreaticcancercellstreated Absenceofanymetastatictumors;controlstreatedwitha2ora3integrin
withb1integrinsiRNA74 subunitsiRNAdisplayedmetastatictumors
Conditionaldeletionofb1integrinfrompancreatic Lossofb1expressioninducedincreasedtumorcellemboliinthelymphatic
bcellscrossedwithRip1Tag2mice68 vasculaturebutnometastasisformation;similarly,tailveininjectionsofb
tumorcellslackingb1integrinexpressiondidnotformmetastases
Ras-myctransformedb1nullEScellsinjected Reducednumbersandsizeofmetastaticfociinthelung
sub-cutaneously79
a5b1integrin HT-29coloncancercellsexpressingexogenousa5 Significantlyreducedlungandextrapulmonarymetastases
integrininjectedintravenously85
LewisLungCarcinomacellsexpressinga5shRNAinjected Fewerlungtumors
intotailvein83
reports that a5 depletion reduced lung colonization in a tail vein model.91 Fibroblasts lacking β3 integrin expression have signific-
injection model.83 However, when a5 and β1 are analyzed antlyfastermigrationspeedsandreducedmigrationpersistencein
separately, the findings are conflicting. Exogenous a5 expression a 3D CDM assay.59 It is difficult to estimate to what degree the
was shown to significantly reduce lung and extrapulmonary usage of different cell types has contributed to the disparate
metastasesarisingfrominjectedcoloncancercells.85Upregulation findings.Forinstance,thedifferentialexpressionofotherintegrin
of β3 expression increased bone and lung metastases,76,81 yet receptors in different cell types may have influenced the invasion
decreased lung tumor formation in an invasive melanoma and metastasis behavior. In particular, it has been suggested that
www.landesbioscience.com CellAdhesion&Migration 429
cross-talkbetweenrecyclinga5β1andaVβ3receptorsmaybean and at the metastatic niches that characterize different tumors.
importantmechanismofregulatingmigrationpersistence.92These Such tailored 3D models to map the journey taken by
datahighlighttheneedfordirectcomparisonbetween3Dculture metastasizing cancer cells are still lacking for interactions with
models and in vivo metastasis models for validation. the vasculature, host immune responses and other cell-cell
interactions, and can generally mimic only short-term (, 1
Conclusion week) conditions.93 But even with these limitations, 3D models
are essential for gaining a basic understanding of cell migration
Our comparison of the effects of a handful of studies that have and invasion through tissue and will ultimately lead to novel
investigated adhesion proteins in 2D vs. 3D culture models with cancer therapies. Invasion away from the primary tumor site
in vivo measurements of metastasis has revealed a complex fundamentally underpins metastatic progression, and treatments
picture. For instance, it is notable that without exception, the that target invasion therefore hold the promise of significantly
depletion of the adhesion molecules summarized in Table 1 improving survival rates for most cancers.
resultedindecreasedmigrationpersistencein3Dmodelsbutnot
always in reduced metastasis formation in vivo (Table 2). To Acknowledgments
accurately predict in vivo invasion behavior, 3D model systems Work in the authors’ labs is supported by National Health and
are required that faithfully replicate the physical characteristics of MedicalResearchCouncilofAustraliagrant632515(G.M.O.),
the environments encountered by transmigrating cancer cells. In NSWCancerCouncilgrantRG12-06(G.M.O.andB.F.),and
particular it will be critical to develop model systems mimicking grants from the German Science Foundation (D.F.G. and
the interstitial tissue organization both at the primary tumor site B.F.).
References 10. WolfK,MüllerR,BorgmannS,BröckerEB,FriedlP. 19. Sabeh F, Shimizu-Hirota R, Weiss SJ. Protease-
Amoeboid shape change and contact guidance: T- dependent versus -independent cancer cell invasion
1. HanahanD,WeinbergRA.Thehallmarksofcancer.
lymphocyte crawling through fibrillar collagen is programs: three-dimensional amoeboid movement
Cell2000;100:57-70;PMID:10647931;http://dx.doi.
independentofmatrixremodelingbyMMPsandother revisited. J Cell Biol 2009; 185:11-9; PMID:
org/10.1016/S0092-8674(00)81683-9
proteases.Blood2003;102:3262-9;PMID:12855577; 19332889;http://dx.doi.org/10.1083/jcb.200807195
2. GritsenkoPG,IlinaO,FriedlP.Interstitialguidanceof
http://dx.doi.org/10.1182/blood-2002-12-3791 20. GeerDJ,SwartzDD,AndreadisST.Fibrinpromotes
cancer invasion. J Pathol 2012; 226:185-99; PMID:
11. Cukierman E, Pankov R, Stevens DR, Yamada KM. migration in a three-dimensional in vitro model of
22006671;http://dx.doi.org/10.1002/path.3031
Taking cell-matrix adhesions to the third dimension. wound regeneration. Tissue Eng 2002; 8:787-98;
3. Kulesa PM, Bailey CM, Kasemeier-Kulesa JC,
Science2001;294:1708-12;PMID:11721053;http:// PMID:12459057; http://dx.doi.org/10.1089/
McLennanR.Cranialneuralcrestmigration:newrules
dx.doi.org/10.1126/science.1064829 10763270260424141
foranoldroad.DevBiol2010;344:543-54;PMID:
12. HakkinenKM,HarunagaJS,DoyleAD,YamadaKM. 21. BrowerJ,BlumbergS,CarrollE,PastarI,BremH,Chen
20399765;http://dx.doi.org/10.1016/j.ydbio.2010.04.
Directcomparisonsofthemorphology,migration,cell W.Mesenchymalstemcelltherapyanddeliverysystems
010
adhesions,andactincytoskeletonoffibroblastsinfour innonhealingwounds.AdvSkinWoundCare2011;24:
4. Cameron DB, Kasai K, Jiang Y, Hu T, Saeki Y,
different three-dimensional extracellular matrices. 524-32, quiz 533-4; PMID:22015751; http://dx.doi.
KomuroH.Fourdistinctphasesofbasket/stellatecell
Tissue Eng Part A 2011; 17:713-24; PMID: org/10.1097/01.ASW.0000407648.89961.a6
migration after entering their final destination (the
20929283; http://dx.doi.org/10.1089/ten.tea.2010. 22. Sanz-MorenoV,GadeaG,AhnJ,PatersonH,MarraP,
molecularlayer)inthedevelopingcerebellum.DevBiol
0273 PinnerS,etal.Racactivationandinactivationcontrol
2009; 332:309-24; PMID:19500566; http://dx.doi.
13. ElsdaleT,BardJ.Collagensubstrataforstudiesoncell plasticityoftumorcellmovement.Cell2008;135:510-
org/10.1016/j.ydbio.2009.05.575
behavior. J Cell Biol 1972; 54:626-37; PMID: 23;PMID:18984162;http://dx.doi.org/10.1016/j.cell.
5. Britto JM, Tait KJ, Johnston LA, Hammond VE,
4339818;http://dx.doi.org/10.1083/jcb.54.3.626 2008.09.043
KalloniatisM,TanSS.Alteredspeedsandtrajectories 14. Wetzels RH, Holland R, van Haelst UJ, Lane EB, 23. LeesJG,BachCT,O’NeillGM.Interiordecoration:
of neurons migrating in the ventricular and subven-
Leigh IM, Ramaekers FC. Detection of basement tropomyosininactindynamicsandcellmigration.Cell
tricular zones of the reeler neocortex. Cereb Cortex
membrane components and basal cell keratin 14 in AdhMigr2011;5:181-6;PMID:21173575;http://dx.
2011; 21:1018-27; PMID:20847150; http://dx.doi.
noninvasiveandinvasivecarcinomasofthebreast.AmJ doi.org/10.4161/cam.5.2.14438
org/10.1093/cercor/bhq168
Pathol1989;134:571-9;PMID:2466404 24. Kotlarchyk MA, Shreim SG, Alvarez-Elizondo MB,
6. SimóS,JossinY,CooperJA.Cullin5regulatescortical
15. Levental KR,YuH,Kass L,LakinsJN, EgebladM, EstradaLC,SinghR,ValdevitL,etal.Concentration
layering by modulating the speed and duration of
Erler JT, et al. Matrix crosslinking forces tumor independentmodulation oflocal micromechanicsina
Dab1-dependentneuronalmigration.JNeurosci2010;
progression by enhancing integrin signaling. Cell fibringel.PLoSOne2011;6:e20201;PMID:21629793;
30:5668-76; PMID:20410119; http://dx.doi.org/10.
2009; 139:891-906; PMID:19931152; http://dx.doi. http://dx.doi.org/10.1371/journal.pone.0020201
1523/JNEUROSCI.0035-10.2010
org/10.1016/j.cell.2009.10.027 25. ZhuYK,UminoT,LiuXD,WangHJ,RombergerDJ,
7. Furnari FB, Fenton T, Bachoo RM, Mukasa A,
16. Shao ZM, Nguyen M, Barsky SH. Human breast SpurzemJR,etal.Effectofinitialcollagenconcentra-
Stommel JM, Stegh A, et al. Malignant astrocytic
carcinoma desmoplasia is PDGF initiated. Oncogene tiononfibroblastmediatedcontractionofcollagengels.
glioma: genetics, biology, and paths to treatment.
2000; 19:4337-45; PMID:10980609; http://dx.doi. Chest2000;117(Suppl1):234S-5S;PMID:10843925;
Genes Dev 2007; 21:2683-710; PMID:17974913;
org/10.1038/sj.onc.1203785 http://dx.doi.org/10.1378/chest.117.5_suppl_1.234S
http://dx.doi.org/10.1101/gad.1596707
17. WalkerRA.Thecomplexitiesofbreastcancerdesmo- 26. Miron-MendozaM,SeemannJ,GrinnellF.Thediffer-
8. SinghMK,IzumchenkoE,Klein-SzantoAJ,Egleston
plasia. Breast Cancer Res 2001; 3:143-5; PMID: entialregulationofcellmotileactivitythroughmatrix
BL, Wolfson M, Golemis EA. Enhanced genetic
11305947;http://dx.doi.org/10.1186/bcr287 stiffness and porosity in three dimensional collagen
instabilityanddasatinibsensitivityinmammarytumor
cellslackingNEDD9.CancerRes2010;70:8907-16; 18. Yamada KM, Pankov R, Cukierman E. Dimensions matrices. Biomaterials 2010; 31:6425-35; PMID:
PMID:20940402; http://dx.doi.org/10.1158/0008- anddynamicsinintegrinfunction.BrazJMedBiolRes 20537378; http://dx.doi.org/10.1016/j.biomaterials.
5472.CAN-10-0353 2003;36:959-66;PMID:12886449;http://dx.doi.org/ 2010.04.064
10.1590/S0100-879X2003000800001
9. Chen CS, Mrksich M, Huang S, Whitesides GM,
Ingber DE. Geometric control of cell life and death.
Science1997;276:1425-8;PMID:9162012;http://dx.
doi.org/10.1126/science.276.5317.1425
430 CellAdhesion&Migration Volume6Issue5
27. YangYL,LeoneLM,KaufmanLJ.Elasticmoduliof 41. McGuirePG,SeedsNW.Theinteractionofplasmino- 55. BakerEL,SrivastavaJ,YuD,BonnecazeRT,Zaman
collagen gels can be predicted from two-dimensional genactivatorwithareconstitutedbasementmembrane MH.Cancercellmigration:integratedrolesofmatrix
confocal microscopy. Biophys J 2009; 97:2051-60; matrix and extracellular macromolecules produced by mechanics and transforming potential. PLoS One
PMID:19804737; http://dx.doi.org/10.1016/j.bpj. culturedepithelialcells.JCellBiochem1989;40:215- 2011; 6:e20355; PMID:21647371; http://dx.doi.org/
2009.07.035 27; PMID:2504731; http://dx.doi.org/10.1002/jcb. 10.1371/journal.pone.0020355
28. GobeauxF,MosserG,AngloA,PanineP,DavidsonP, 240400210 56. Kim HD, Guo TW, Wu AP, Wells A, Gertler FB,
Giraud-Guille MM, et al. Fibrillogenesis in dense 42. Li ML, Aggeler J, Farson DA, Hatier C, Hassell J, Lauffenburger DA. Epidermal growth factor-induced
collagensolutions:aphysicochemicalstudy.JMolBiol Bissell MJ. Influence of a reconstituted basement enhancement of glioblastoma cell migration in 3D
2008; 376:1509-22; PMID:18234220; http://dx.doi. membrane and its components on casein gene arisesfromanintrinsicincreaseinspeedbutanextrinsic
org/10.1016/j.jmb.2007.12.047 expressionandsecretioninmousemammaryepithelial matrix- and proteolysis-dependent increase in persist-
29. MickelW,MünsterS,JawerthLM,VaderDA,Weitz cells. Proc Natl Acad Sci U S A 1987; 84:136-40; ence. Mol Biol Cell 2008; 19:4249-59; PMID:
DA,SheppardAP,etal.Robustporesizeanalysisof PMID:3467345; http://dx.doi.org/10.1073/pnas.84.1. 18632979; http://dx.doi.org/10.1091/mbc.E08-05-
filamentousnetworksfromthree-dimensionalconfocal 136 0501
microscopy. Biophys J 2008; 95:6072-80; PMID: 43. LiW,MachuleD,GaoC,DenBestenPK.Growthof 57. Indra I, Undyala V, Kandow C, Thirumurthi U,
18835899; http://dx.doi.org/10.1529/biophysj.108. ameloblast-lineagecellsinathree-dimensionalMatrigel Dembo M, Beningo KA. An in vitro correlation of
135939 environment.EurJOralSci2006;114(Suppl1):159- mechanical forces and metastatic capacity. Phys Biol
30. McPhersonJM,WallaceDG,SawamuraSJ,ContiA, 63,discussion164-5,380-1;PMID:16674679;http:// 2011; 8:015015; PMID:21301068; http://dx.doi.org/
CondellRA,WadeS,etal.Collagenfibrillogenesisin dx.doi.org/10.1111/j.1600-0722.2006.00308.x 10.1088/1478-3975/8/1/015015
vitro:acharacterizationoffibrilqualityasafunctionof 44. KubotaY,KleinmanHK,MartinGR,LawleyTJ.Role 58. DiMilla PA, Stone JA, Quinn JA, Albelda SM,
assembly conditions. Coll Relat Res 1985; 5:119-35; of laminin and basement membrane in the morpho- Lauffenburger DA. Maximal migration of human
PMID:3924470; http://dx.doi.org/10.1016/S0174- logical differentiation of human endothelial cells into smooth muscle cells on fibronectin and type IV
173X(85)80034-0 capillary-likestructures.JCellBiol1988;107:1589-98; collagenoccursatanintermediateattachmentstrength.
31. Zeugolis DI, Paul RG, Attenburrow G. Factors PMID:3049626; http://dx.doi.org/10.1083/jcb.107.4. JCellBiol1993;122:729-37;PMID:8335696;http://
influencing the properties of reconstituted collagen 1589 dx.doi.org/10.1083/jcb.122.3.729
fiberspriortoself-assembly:animalspeciesandcollagen 45. Carey DJ, Todd MS, Rafferty CM. Schwann cell 59. WorthDC,Hodivala-DilkeK,RobinsonSD,KingSJ,
extractionmethod.JBiomedMaterResA2008;86: myelination:inductionbyexogenousbasementmem- MortonPE,GertlerFB,etal.Alphavbeta3integrin
892-904; PMID:18041730; http://dx.doi.org/10. brane-likeextracellularmatrix.JCellBiol1986;102: spatiallyregulatesVASPandRIAMtocontroladhesion
1002/jbm.a.31694 2254-63; PMID:3086325; http://dx.doi.org/10.1083/ dynamicsandmigration.JCellBiol2010;189:369-83;
32. RaubCB,SureshV,KrasievaT,LyubovitskyJ,Mih jcb.102.6.2254 PMID:20404115; http://dx.doi.org/10.1083/jcb.
JD, Putnam AJ, et al. Noninvasive assessment of 46. PorterKR,VanameeP.Observationsontheformation 200912014
collagengelmicrostructureandmechanicsusingmulti- of connective tissue fibers. Proc Soc Exp Biol Med 60. ZhongJ,BaquiranJB,BonakdarN,LeesJ,ChingYW,
photon microscopy. Biophys J 2007; 92:2212-22; 1949;71:513-6;PMID:18139799 PugachevaE,etal.NEDD9stabilizesfocaladhesions,
PMID:17172303; http://dx.doi.org/10.1529/biophysj. 47. Hedman K, Vaheri A, Wartiovaara J. External increases binding to the extra-cellular matrix and
106.097998 fibronectinofculturedhumanfibroblastsispredomi- differentially effects 2D versus 3D cell migration.
33. Bannister DW,BurnsAB.Pepsin treatment ofavian nantlyamatrixprotein.JCellBiol1978;76:748-60; PLoSOne2012;7:e35058;PMID:22509381;http://
skincollagen.Effectsonsolubility,subunitcomposition PMID:344328; http://dx.doi.org/10.1083/jcb.76.3. dx.doi.org/10.1371/journal.pone.0035058
andaggregationproperties.BiochemJ1972;129:677- 748 61. Sanz-Moreno V, Marshall CJ. The plasticity of
81;PMID:4572795 48. VaheriA,KurkinenM,LehtoVP,LinderE,TimplR. cytoskeletaldynamicsunderlyingneoplasticcellmigra-
34. SnowdenJM,SwannDA.Theformationandthermal Codistribution of pericellular matrix proteins in tion. Curr Opin Cell Biol 2010; 22:690-6; PMID:
stabilityofinvitroassembledfibrilsfromacid-soluble cultured fibroblasts and loss in transformation: fibro- 20829016; http://dx.doi.org/10.1016/j.ceb.2010.08.
and pepsin-treated collagens. Biochim Biophys Acta nectin and procollagen. Proc Natl Acad Sci U S A 020
1979;580:372-81;PMID:42446;http://dx.doi.org/10. 1978;75:4944-8;PMID:217006;http://dx.doi.org/10. 62. MierkeCT,FreyB,FellnerM,HerrmannM,FabryB.
1016/0005-2795(79)90149-1 1073/pnas.75.10.4944 Integrin a5β1 facilitates cancer cell invasion through
35. Forgacs G, Newman SA, Hinner B, Maier CW, 49. Hedman K, Kurkinen M, Alitalo K, Vaheri A, enhancedcontractileforces.JCellSci2011;124:369-
SackmannE.Assemblyofcollagenmatricesasaphase Johansson S, Höök M. Isolation of the pericellular 83; PMID:21224397; http://dx.doi.org/10.1242/jcs.
transitionrevealedbystructuralandrheologicstudies. matrixofhumanfibroblastcultures.JCellBiol1979; 071985
BiophysJ2003;84:1272-80;PMID:12547807;http:// 81:83-91; PMID:383722; http://dx.doi.org/10.1083/ 63. GuckJ,SchinkingerS,LincolnB,WottawahF,Ebert
dx.doi.org/10.1016/S0006-3495(03)74942-X jcb.81.1.83 S, Romeyke M, et al. Optical deformability as an
36. NotbohmH,MoslerS,MüllerPK,BrinckmannJ.In 50. Narayanan K, Leck KJ, Gao S, Wan AC. Three- inherentcellmarkerfortestingmalignanttransforma-
vitroformationandaggregationofheterotypiccollagen dimensional reconstituted extracellular matrix scaf- tionandmetastaticcompetence.BiophysJ2005;88:
Iand IIIfibrils.Int JBiolMacromol 1993;15:299- folds for tissue engineering. Biomaterials 2009; 30: 3689-98; PMID:15722433; http://dx.doi.org/10.
304; PMID:8251445; http://dx.doi.org/10.1016/ 4309-17; PMID:19477508; http://dx.doi.org/10. 1529/biophysj.104.045476
0141-8130(93)90030-P 1016/j.biomaterials.2009.04.049 64. RemmerbachTW,WottawahF,DietrichJ,LincolnB,
37. Timpl R, Martin GR, Bruckner P, Wick G, 51. PankovR,EndoY,Even-RamS,ArakiM,ClarkK, Wittekind C, Guck J. Oral cancer diagnosis by
Wiedemann H.Natureofthecollagenousproteinin Cukierman E, et al. A Rac switch regulates random mechanicalphenotyping.CancerRes2009;69:1728-
atumorbasementmembrane.EurJBiochem1978;84: versusdirectionallypersistentcellmigration.JCellBiol 32; PMID:19223529; http://dx.doi.org/10.1158/
43-52; PMID:648517; http://dx.doi.org/10.1111/j. 2005; 170:793-802; PMID:16129786; http://dx.doi. 0008-5472.CAN-08-4073
1432-1033.1978.tb12139.x org/10.1083/jcb.200503152 65. KochTM,MünsterS,BonakdarN,ButlerJP,FabryB.
38. OrkinRW,GehronP,McGoodwinEB,MartinGR, 52. Fraley SI, Feng Y, Krishnamurthy R, Kim DH, 3DTractionforcesincancercellinvasion.PLoSOne
ValentineT,SwarmR.Amurinetumorproducinga Celedon A, Longmore GD, et al. A distinctive role 2012; 7:e33476; PMID:22479403; http://dx.doi.org/
matrixofbasementmembrane.JExpMed1977;145: for focal adhesion proteins in three-dimensional cell 10.1371/journal.pone.0033476
204-20; PMID:830788; http://dx.doi.org/10.1084/ motility. Nat Cell Biol 2010; 12:598-604; PMID: 66. Kraning-Rush CM, Califano JP, Reinhart-King CA.
jem.145.1.204 20473295;http://dx.doi.org/10.1038/ncb2062 Cellular traction stresses increase with increasing
39. Kleinman HK. Preparation of basement membrane 53. Kubow KE, Horwitz AR. Reducing background metastatic potential. PLoS One 2012; 7:e32572;
components from ehs tumors. Curr Protoc Cell Biol fluorescence reveals adhesions in 3D matrices. Nat PMID:22389710; http://dx.doi.org/10.1371/journal.
2001;Chapter10:Unit102. Cell Biol 2011; 13:3-5, author reply 5-7; PMID: pone.0032572
40. KleinmanHK,McGarveyML,LiottaLA,RobeyPG, 21173800;http://dx.doi.org/10.1038/ncb0111-3
TryggvasonK,MartinGR.Isolationandcharacteriza- 54. Harunaga JS, Yamada KM. Cell-matrix adhesions in
tion of type IV procollagen, laminin, and heparan 3D. Matrix Biol 2011; 30:363-8; PMID:21723391;
sulfate proteoglycan from the EHS sarcoma. http://dx.doi.org/10.1016/j.matbio.2011.06.001
Biochemistry 1982; 21:6188-93; PMID:6217835;
http://dx.doi.org/10.1021/bi00267a025
www.landesbioscience.com CellAdhesion&Migration 431
67. Izumchenko E, Singh MK, Plotnikova OV, 76. SloanEK,PouliotN,StanleyKL,ChiaJ,MoseleyJM, 86. McLean GW,CarragherNO,AvizienyteE,EvansJ,
Tikhmyanova N, Little JL, Serebriiskii IG, et al. Hards DK, et al. Tumor-specific expression of BruntonVG,FrameMC.Theroleoffocal-adhesion
NEDD9 promotes oncogenic signaling in mammary alphavbeta3integrinpromotesspontaneousmetastasis kinaseincancer—anewtherapeuticopportunity.Nat
tumor development. Cancer Res2009; 69:7198-206; ofbreastcancertobone.BreastCancerRes2006;8: RevCancer2005;5:505-15;PMID:16069815;http://
PMID:19738060; http://dx.doi.org/10.1158/0008- R20; PMID:16608535; http://dx.doi.org/10.1186/ dx.doi.org/10.1038/nrc1647
5472.CAN-09-0795 bcr1398 87. Mierke CT, Kollmannsberger P,Zitterbart DP, Diez
68. Kren A, Baeriswyl V, Lehembre F, Wunderlin C, 77. CabodiS,TinnirelloA,BisaroB,TornilloG,delPilar G, Koch TM, Marg S, et al. Vinculin facilitates cell
StrittmatterK,AntoniadisH,etal.Increasedtumorcell Camacho-Leal M, Forni G, et al. p130Cas is an invasion into three-dimensional collagen matrices. J
disseminationandcellularsenescenceintheabsenceof essentialtransducerelementinErbB2transformation. Biol Chem 2010; 285:13121-30; PMID:20181946;
beta1-integrin function. EMBO J2007; 26:2832-42; FASEB J 2010; 24:3796-808; PMID:20505116; http://dx.doi.org/10.1074/jbc.M109.087171
PMID:17541405; http://dx.doi.org/10.1038/sj.emboj. http://dx.doi.org/10.1096/fj.10-157347 88. O’NeillGM,SeoS,SerebriiskiiIG,LessinSR,Golemis
7601738 78. BrábekJ,ConstancioSS,SiesserPF,ShinNY,PozziA, EA. A new central scaffold for metastasis: parsing
69. Lahlou H, Sanguin-Gendreau V, Frame MC, Muller HanksSK.Crk-associatedsubstratetyrosinephosphor- HEF1/Cas-L/NEDD9. Cancer Res 2007; 67:8975-9;
WJ.Focaladhesionkinasecontributestoproliferative ylationsitesarecriticalforinvasionandmetastasisof PMID:17908996; http://dx.doi.org/10.1158/0008-
potential of ErbB2 mammary tumour cells but is SRC-transformedcells.MolCancerRes2005;3:307- 5472.CAN-07-1328
dispensableforErbB2mammarytumourinductionin 15; PMID:15972849; http://dx.doi.org/10.1158/ 89. Paszek MJ, Zahir N, Johnson KR, Lakins JN,
vivo. Breast Cancer Res 2012; 14:R36; PMID: 1541-7786.MCR-05-0015 RozenbergGI,GefenA,etal.Tensionalhomeostasis
22373082;http://dx.doi.org/10.1186/bcr3131 79. BrakebuschC,WennerbergK,KrellHW,WeidleUH, and the malignant phenotype. Cancer Cell 2005; 8:
70. LahlouH,Sanguin-GendreauV,ZuoD,CardiffRD, SallmyrA,JohanssonS,etal.Beta1integrinpromotes 241-54; PMID:16169468; http://dx.doi.org/10.1016/
McLeanGW,FrameMC,etal.Mammaryepithelial- butisnotessentialformetastasisofras-myctransformed j.ccr.2005.08.010
specificdisruptionofthefocaladhesionkinaseblocks fibroblasts. Oncogene 1999; 18:3852-61; PMID: 90. ZamanMH,TrapaniLM,SieminskiAL,MackellarD,
mammarytumorprogression.ProcNatlAcadSciUSA 10445848;http://dx.doi.org/10.1038/sj.onc.1202770 GongH,KammRD,etal.Migrationoftumorcellsin
2007; 104:20302-7; PMID:18056629; http://dx.doi. 80. Rodríguez Fernández JL, Geiger B, Salomon D, 3Dmatricesisgovernedbymatrixstiffnessalongwith
org/10.1073/pnas.0710091104 Sabanay I, Zöller M, Ben-Ze’ev A. Suppression of cell-matrixadhesionandproteolysis.ProcNatlAcadSci
71. Provenzano PP, InmanDR,EliceiriKW,Beggs HE, tumorigenicity in transformed cells after transfection USA2006;103:10889-94;PMID:16832052;http://
Keely PJ. Mammary epithelial-specific disruption of with vinculin cDNA. J Cell Biol 1992; 119:427-38; dx.doi.org/10.1073/pnas.0604460103
focal adhesion kinase retards tumor formation and PMID:1400584; http://dx.doi.org/10.1083/jcb.119.2. 91. DanenEH,vanKraatsAA,CornelissenIM,RuiterDJ,
metastasisinatransgenicmousemodelofhumanbreast 427 van Muijen GN. Integrin beta 3 cDNA transfection
cancer. Am J Pathol 2008; 173:1551-65; PMID: 81. Felding-Habermann B, O’Toole TE, Smith JW, intoahighlymetastaticalphavbeta3-negativehuman
18845837; http://dx.doi.org/10.2353/ajpath.2008. Fransvea E, Ruggeri ZM, Ginsberg MH, et al. melanomacelllineinhibitsinvasionandexperimental
080308 Integrinactivationcontrolsmetastasisinhumanbreast metastasis.BiochemBiophysResCommun1996;226:
72. Huck L, Pontier SM, Zuo DM, Muller WJ. beta1- cancer.ProcNatlAcadSciUSA2001;98:1853-8; 75-81; PMID:8806594; http://dx.doi.org/10.1006/
integrin is dispensable for the induction of ErbB2 PMID:11172040; http://dx.doi.org/10.1073/pnas.98. bbrc.1996.1313
mammary tumors but plays a critical role in the 4.1853 92. WhiteDP,CaswellPT,NormanJC.alphavbeta3and
metastaticphaseoftumorprogression.ProcNatlAcad 82. Pylayeva Y, Gillen KM, Gerald W, Beggs HE, alpha5beta1integrinrecyclingpathwaysdictatedown-
Sci U S A 2010; 107:15559-64; PMID:20713705; ReichardtLF,GiancottiFG.Ras-andPI3K-dependent streamRhokinasesignalingtoregulatepersistentcell
http://dx.doi.org/10.1073/pnas.1003034107 breasttumorigenesisinmiceandhumansrequiresfocal migration. J Cell Biol 2007; 177:515-25; PMID:
73. DuxburyMS,ItoH,ZinnerMJ,AshleySW,Whang adhesionkinasesignaling.JClinInvest2009;119:252- 17485491;http://dx.doi.org/10.1083/jcb.200609004
EE. Focal adhesion kinase gene silencing promotes 66;PMID:19147981 93. YamadaKM,CukiermanE.Modelingtissuemorpho-
anoikisandsuppressesmetastasisofhumanpancreatic 83. RomanJ,RitzenthalerJD,Roser-PageS,SunX,HanS. genesis and cancer in 3D. Cell 2007; 130:601-10;
adenocarcinoma cells. Surgery 2004; 135:555-62; alpha5beta1-integrin expression is essential for tumor PMID:17719539; http://dx.doi.org/10.1016/j.cell.
PMID:15118593; http://dx.doi.org/10.1016/j.surg. progressioninexperimentallungcancer.AmJRespir 2007.08.006
2003.10.017 Cell Mol Biol 2010; 43:684-91; PMID:20081050; 94. DeakinNO,TurnerCE.Distinctrolesforpaxillinand
74. Grzesiak JJ, Tran Cao HS, Burton DW, Kaushal S, http://dx.doi.org/10.1165/rcmb.2009-0375OC Hic-5 in regulating breast cancer cell morphology,
Vargas F, Clopton P, et al. Knockdown of the 84. Sakamoto S, McCann RO, Dhir R, Kyprianou N. invasion,andmetastasis.MolBiolCell2011;22:327-
β(1) integrin subunit reduces primary tumor growth Talin1 promotes tumor invasion and metastasis via 41;PMID:21148292;http://dx.doi.org/10.1091/mbc.
andinhibitspancreaticcancermetastasis.IntJCancer focaladhesionsignalingandanoikisresistance.Cancer E10-09-0790
2011; 129:2905-15; PMID:21491421; http://dx.doi. Res 2010; 70:1885-95; PMID:20160039; http://dx.
org/10.1002/ijc.25942 doi.org/10.1158/0008-5472.CAN-09-2833
75. HsuMY,ShihDT,MeierFE,VanBelleP,HsuJY, 85. Schirner M, Herzberg F, Schmidt R, Streit M,
Elder DE, et al. Adenoviral gene transfer of beta3 SchöningM,HummelM,etal.Integrinalpha5beta1:
integrin subunit induces conversion from radial to apotentinhibitorofexperimentallungmetastasis.Clin
verticalgrowthphaseinprimaryhumanmelanoma.Am Exp Metastasis 1998; 16:427-35; PMID:10091938;
JPathol1998;153:1435-42;PMID:9811334;http:// http://dx.doi.org/10.1023/A:1006581424490
dx.doi.org/10.1016/S0002-9440(10)65730-6
432 CellAdhesion&Migration Volume6Issue5