How does invasion influence tumor growth

Blood 24 — J Clin Invest 11 — Jakobsson L et al Endothelial cells dynamically compete for the tip cell position during angiogenic sprouting. Nat Cell Biol 12 10 — Hellstrom M et al Dll4 signalling through Notch1 regulates formation of tip cells during angiogenesis.

PubMed Google Scholar. Harrington LS et al Regulation of multiple angiogenic pathways by Dll4 and Notch in human umbilical vein endothelial cells. Microvasc Res 75 2 — J Angiogenes Res 2 1 J Cell Biol 6 — Fantin A et al NRP1 acts cell autonomously in endothelium to promote tip cell function during sprouting angiogenesis. Blood 12 — Blood 19 — Phng LK et al Nrarp coordinates endothelial Notch and Wnt signaling to control vessel density in angiogenesis.

Dev Cell 16 1 — Herwig L et al Distinct cellular mechanisms of blood vessel fusion in the zebrafish embryo. Curr Biol 21 22 — Kochhan E et al Blood flow changes coincide with cellular rearrangements during blood vessel pruning in zebrafish embryos. PLoS One 8 10 :e Lenard A et al Endothelial cell self-fusion during vascular pruning. PLoS Biol 13 4 :e Lenard A et al In vivo analysis reveals a highly stereotypic morphogenetic pathway of vascular anastomosis.

Dev Cell 25 5 — Development 19 — Blum Y et al Complex cell rearrangements during intersegmental vessel sprouting and vessel fusion in the zebrafish embryo. Dev Biol 2 — Betz C et al Cell behaviors and dynamics during angiogenesis. Development 13 — Patan S et al Intussusceptive microvascular growth: a common alternative to capillary sprouting. Arch Histol Cytol 55 Suppl — Anat Rec 1 — Development 14 — Wilting J et al VEGF induces proliferation of vascular endothelial cells and expression of flk-1 without affecting lymphatic vessels of chorioallantoic membrane.

Dev Biol 1 — Crivellato E et al Recombinant human erythropoietin induces intussusceptive microvascular growth in vivo. Leukemia 18 2 — Ribatti D et al Microvascular density, vascular endothelial growth factor immunoreactivity in tumor cells, vessel diameter and intussusceptive microvascular growth in primary melanoma.

Oncol Rep 14 1 — Nico B et al Intussusceptive microvascular growth in human glioma. Clin Exp Med 10 2 — Patan S, Munn LL, Jain RK Intussusceptive microvascular growth in a human colon adenocarcinoma xenograft: a novel mechanism of tumor angiogenesis.

Microvasc Res 51 2 — Djonov V et al MMP cellular localization of a novel metalloproteinase within normal breast tissue and mammary gland tumours. J Pathol 2 — Risau W et al Vasculogenesis and angiogenesis in embryonic-stem-cell-derived embryoid bodies. Development 3 — Risau W, Lemmon V Changes in the vascular extracellular matrix during embryonic vasculogenesis and angiogenesis. Choi K Hemangioblast development and regulation.

Biochem Cell Biol 76 6 — Asahara T et al Isolation of putative progenitor endothelial cells for angiogenesis. Science — Kioi M et al Inhibition of vasculogenesis, but not angiogenesis, prevents the recurrence of glioblastoma after irradiation in mice.

J Clin Invest 3 — Cancer Lett 1 — Chopra H et al Insights into endothelial progenitor cells: origin, classification, potentials, and prospects. Stem Cells Int Circ Res 2 — Circ Res 97 4 — Shin JW et al Isolation of endothelial progenitor cells from cord blood and induction of differentiation by ex vivo expansion.

Yonsei Med J 46 2 — Urbich C, Dimmeler S Endothelial progenitor cells: characterization and role in vascular biology. Circ Res 95 4 — Reale A et al Functional and biological role of endothelial precursor cells in tumour progression: a new potential therapeutic target in haematological malignancies. Asahara T et al VEGF contributes to postnatal neovascularization by mobilizing bone marrow-derived endothelial progenitor cells.

EMBO J 18 14 — Hattori K et al Vascular endothelial growth factor and angiopoietin-1 stimulate postnatal hematopoiesis by recruitment of vasculogenic and hematopoietic stem cells. J Exp Med 9 — Kopp HG, Ramos CA, Rafii S Contribution of endothelial progenitors and proangiogenic hematopoietic cells to vascularization of tumor and ischemic tissue.

Curr Opin Hematol 13 3 — Chang EI et al Hypoxia, hormones, and endothelial progenitor cells in hemangioma. Lymphat Res Biol 5 4 — Spring H et al Chemokines direct endothelial progenitors into tumor neovessels. FEBS Lett 15 — Maniotis AJ et al Vascular channel formation by human melanoma cells in vivo and in vitro: vasculogenic mimicry. Am J Pathol 3 — Ricci-Vitiani L et al Tumour vascularization via endothelial differentiation of glioblastoma stem-like cells. Upile T et al Vascular mimicry in cultured head and neck tumour cell lines.

Head Neck Oncol Williamson SC et al Vasculogenic mimicry in small cell lung cancer. Nat Commun Baeten CI et al Prognostic role of vasculogenic mimicry in colorectal cancer. Dis Colon Rectum 52 12 — Sharma N et al Prostatic tumor cell plasticity involves cooperative interactions of distinct phenotypic subpopulations: role in vasculogenic mimicry. Prostate 50 3 — Fausto N Vasculogenic mimicry in tumors.

Fact or artifact? Am J Pathol 2 Seftor RE et al Tumor cell vasculogenic mimicry: from controversy to therapeutic promise. Am J Pathol 4 — APMIS 7—8 — Transl Oncol 10 4 — Valyi-Nagy K et al Stem cell marker CD is expressed by vasculogenic mimicry-forming uveal melanoma cells in three-dimensional cultures. Mol Vis — Lin AY et al Distinguishing fibrovascular septa from vasculogenic mimicry patterns.

Arch Pathol Lab Med 7 — Free Radic Biol Med 51 4 — Histol Histopathol 32 9 — Li M et al Vasculogenic mimicry: a new prognostic sign of gastric adenocarcinoma.

Pathol Oncol Res 16 2 — Wang R et al Glioblastoma stem-like cells give rise to tumour endothelium. Mei X et al Glioblastoma stem cell differentiation into endothelial cells evidenced through live-cell imaging. Neuro Oncol 19 8 — J Cell Mol Med 13 2 — Alvero AB et al Stem-like ovarian cancer cells can serve as tumor vascular progenitors.

Stem Cells 27 10 — Zhao Y et al Endothelial cell transdifferentiation of human glioma stem progenitor cells in vitro. Brain Res Bull 82 5—6 — Kulla A et al Analysis of the TP53 gene in laser-microdissected glioblastoma vasculature. Acta Neuropathol 4 — Rodriguez FJ et al Neoplastic cells are a rare component in human glioblastoma microvasculature. Oncotarget 3 1 — De Palma M et al Tie2 identifies a hematopoietic lineage of proangiogenic monocytes required for tumor vessel formation and a mesenchymal population of pericyte progenitors.

Cancer Cell 8 3 — Cheng L et al Glioblastoma stem cells generate vascular pericytes to support vessel function and tumor growth. Cell 1 — Curr Opin Genet Dev 15 1 — McDonald DM, Baluk P Imaging of angiogenesis in inflamed airways and tumors: newly formed blood vessels are not alike and may be wildly abnormal: Parker B.

Francis lecture. Chest 6 Suppl S—S. Kimura H et al Fluctuations in red cell flux in tumor microvessels can lead to transient hypoxia and reoxygenation in tumor parenchyma. Cancer Res 56 23 — Cancer Res 64 17 — Hashizume H et al Openings between defective endothelial cells explain tumor vessel leakiness.

Padera TP et al Pathology: cancer cells compress intratumour vessels. Nature Abramsson A et al Analysis of mural cell recruitment to tumor vessels. Circulation 1 — Morikawa S et al Abnormalities in pericytes on blood vessels and endothelial sprouts in tumors. Baluk P et al Abnormalities of basement membrane on blood vessels and endothelial sprouts in tumors. Am J Pathol 5 — St Croix B et al Genes expressed in human tumor endothelium. Zhang L et al Tumor-derived vascular endothelial growth factor up-regulates angiopoietin-2 in host endothelium and destabilizes host vasculature, supporting angiogenesis in ovarian cancer.

Cancer Res 63 12 — Carson-Walter EB et al Cell surface tumor endothelial markers are conserved in mice and humans. Cancer Res 61 18 — Huang X et al Lymphoma endothelium preferentially expresses Tim-3 and facilitates the progression of lymphoma by mediating immune evasion.

J Exp Med 3 — J Pathol 3 — Roudnicky F et al Endocan is upregulated on tumor vessels in invasive bladder cancer where it mediates VEGF-A-induced angiogenesis. Cancer Res 73 3 — Zhao Q et al Single-cell transcriptome analyses reveal endothelial cell heterogeneity in tumors and changes following antiangiogenic treatment. Cancer Res 78 9 — Buckanovich RJ et al Tumor vascular proteins as biomarkers in ovarian cancer.

J Clin Oncol 25 7 — Zhang L et al IDH mutation status is associated with distinct vascular gene expression signatures in lower-grade gliomas. Neuro Oncol 20 11 — Masiero M et al A core human primary tumor angiogenesis signature identifies the endothelial orphan receptor ELTD1 as a key regulator of angiogenesis.

Cancer Cell 24 2 — Semin Thromb Hemost 33 7 — Maruno M et al Expression of thrombomodulin in astrocytomas of various malignancy and in gliotic and normal brains. J Neurooncol 19 2 — Oncogene 31 3 — Langenkamp E et al Elevated expression of the C-type lectin CD93 in the glioblastoma vasculature regulates cytoskeletal rearrangements that enhance vessel function and reduce host survival.

Cancer Res 75 21 — Lugano R et al CD93 promotes beta1 integrin activation and fibronectin fibrillogenesis during tumor angiogenesis. J Clin Invest 8 — Christian S et al Endosialin Tem1 is a marker of tumor-associated myofibroblasts and tumor vessel-associated mural cells.

Am J Pathol 2 — Oncogene 36 44 — Galvagni F et al Dissecting the CDMultimerin 2 interaction involved in cell adhesion and migration of the activated endothelium. Matrix Biol — Mogler C et al Hepatic stellate cell-expressed endosialin balances fibrogenesis and hepatocyte proliferation during liver damage.

Viski C et al Endosialin-expressing pericytes promote metastatic dissemination. Cancer Res 76 18 — Griffioen AW et al Tumor angiogenesis is accompanied by a decreased inflammatory response of tumor-associated endothelium.

Blood 88 2 — Griffioen AW et al Endothelial intercellular adhesion molecule-1 expression is suppressed in human malignancies: the role of angiogenic factors. Cancer Res 56 5 — Dirkx AE et al Tumor angiogenesis modulates leukocyte-vessel wall interactions in vivo by reducing endothelial adhesion molecule expression. Cancer Res 63 9 — Motz GT et al Tumor endothelium FasL establishes a selective immune barrier promoting tolerance in tumors.

Nat Med 20 6 — Buckanovich RJ et al Endothelin B receptor mediates the endothelial barrier to T cell homing to tumors and disables immune therapy. Nat Med 14 1 — Phoenix TN et al Medulloblastoma genotype dictates blood brain barrier phenotype. Cancer Cell 29 4 — Nat Med 9 6 — Cell 6 — Ferrara N Vascular endothelial growth factor: basic science and clinical progress.

Endocr Rev 25 4 — J Intern Med 2 — EMBO J 20 11 — Adv Cancer Res — Circ Res 6 — Cardiovasc Res 78 2 — Front Oncol J Cell Biol 2 — Hofer E, Schweighofer B Signal transduction induced in endothelial cells by growth factor receptors involved in angiogenesis. Thromb Haemost 97 3 — Nat Med 9 7 — Schomber T et al Placental growth factor-1 attenuates vascular endothelial growth factor-A-dependent tumor angiogenesis during beta cell carcinogenesis.

Cancer Res 67 22 — Cell 3 — Bais C et al PlGF blockade does not inhibit angiogenesis during primary tumor growth. Turner N, Grose R Fibroblast growth factor signalling: from development to cancer. Nat Rev Cancer 10 2 — Wiley Interdiscip Rev Dev Biol 4 3 — Cytokine Growth Factor Rev 16 2 — Compagni A et al Fibroblast growth factors are required for efficient tumor angiogenesis. Cancer Res 60 24 — Yu P et al FGF-dependent metabolic control of vascular development. Sci Transl Med 10 :eaag Physiol Rev 79 4 — Blood 10 — Cytokine Growth Factor Rev 15 4 — Guo P et al Platelet-derived growth factor-B enhances glioma angiogenesis by stimulating vascular endothelial growth factor expression in tumor endothelia and by promoting pericyte recruitment.

Davis S et al Isolation of angiopoietin-1, a ligand for the TIE2 receptor, by secretion-trap expression cloning. Cell 87 7 — Maisonpierre PC et al Angiopoietin-2, a natural antagonist for Tie2 that disrupts in vivo angiogenesis. Springer, Cham, pp 1— Google Scholar. Reiss Y et al Switching of vascular phenotypes within a murine breast cancer model induced by angiopoietin J Pathol 4 — Mol Cancer Res 5 7 — Fiedler U et al Angiopoietin-2 sensitizes endothelial cells to TNF-alpha and has a crucial role in the induction of inflammation.

Nat Med 12 2 — Clin Cancer Res 16 14 — Peterson TE et al Dual inhibition of Ang-2 and VEGF receptors normalizes tumor vasculature and prolongs survival in glioblastoma by altering macrophages. Wu FT et al Efficacy of cotargeting angiopoietin-2 and the VEGF pathway in the adjuvant postsurgical setting for early breast, colorectal, and renal cancers. Cancer Res 76 23 — Lisle JE et al Eph receptors and their ligands: promising molecular biomarkers and therapeutic targets in prostate cancer.

Biochim Biophys Acta 2 — Nat Rev Mol Cell Biol 3 7 — Holder N, Klein R Eph receptors and ephrins: effectors of morphogenesis. Development 10 — Essential mediators of vascular development. Trends Cardiovasc Med 10 5 — Cytokine Growth Factor Rev 15 6 — Oncogene 19 49 — Dong Y et al Downregulation of EphA1 in colorectal carcinomas correlates with invasion and metastasis. Mod Pathol 22 1 — Hafner C et al Loss of EphB6 expression in metastatic melanoma. Int J Oncol 23 6 — Ogawa K et al The ephrin-A1 ligand and its receptor, EphA2, are expressed during tumor neovascularization.

Oncogene 19 52 — Dobrzanski P et al Antiangiogenic and antitumor efficacy of EphA2 receptor antagonist. Growth 30, — Pubmed Abstract Pubmed Full Text. Butler, G. Collier, I. Diffusion of MMPs on the surface of collagen fibrils: the mobile cell surface-collagen substratum interface.

CrossRef Full Text. Robustness analysis and behavior discrimination in enzymatic reaction networks. Membrane-type matrix metalloproteinases 1 and 2 exhibit broad-spectrum proteolytic capacities comparable to many matrix metalloproteinases. English, W. Friedl, P. Proteolytic and non-proteolytic migration of tumour cells and leucocytes.

Gerisch, A. Mathematical modelling of cancer cell invasion of tissue: local and non-local models and the effect of adhesion. Hanahan, D. The hallmarks of cancer. Cell , 57— Hallmarks of cancer: the next generation.

Cell , — Hoshino, D. PLoS Comput. Ichikawa, K. A-Cell: graphical user interface for the construction of biochemical reaction models. Bioinformatics 17, — Karagiannis, E.

A theoretical model of type I collagen proteolysis by matrix metalloproteinase MMP 2 and membrane type 1 MMP in the presence of tissue inhibitor of metalloproteinase 2.

Distinct modes of collagen type I proteolysis by matrix metalloproteinase MMP 2 and membrane type I MMP during the migration of a tip endothelial cell: insights from a computational model. Kessenbrock, K. Matrix metalloproteinases: regulators of the tumor microenvironment. Cell , 52— Kinoh, H. Kleiner, D. Matrix metalloproteinases and metastasis. Cancer Chemother. Lafleur, M. FEBS Lett. Li, X. Molecular dissection of the structural machinery underlying the tissue-invasive activity of membrane type-1 matrix metalloproteinase.

Cell 19, — Protease degradomics: a new challenge for proteomics. Cell Biol. Lowengrub, J. Nonlinear modelling of cancer: bridging the gap between cells and tumours. Nonlinearity 23, R1—R9. Martins, V. Increased invasive behaviour in cutaneous squamous cell carcinoma with loss of basement-membrane type VII collagen.

Nagase, H. Matrix metalloproteinases. Nishida, Y. Cancer Res. New and paradoxical roles of matrix metalloproteinases in the tumor microenvironment. We focused on the 50— kDa fraction for further studies since this fraction contained the majority of the tumor promoting activity.

These results suggest that the size of the heat sensitive factor secreted by astrocytes is greater than 50 kDa. By analyzing the molecular weight of the known astrocyte secretome Table S1 from the literature, the majority of proteins were excluded because their molecular weight is less than 50 kDa.

However, matrix metalloprotease MMP enzymes secreted by astrocytes are in the molecular weight MW range of 50— kDa, and are known to play an important role in tumor invasion and angiogenesis by mediating degradation of the extracellular matrix, which promotes metastasis [21].

We investigated whether MMPs are involved in the astrocyte secretome-induced tumor cell invasion. Our first approach was a chemical biology approach where astrocyte CMs treated separately with broad-spectrum MMP inhibitors ONO, Batimastat and Marimastat were tested on tumor cells in a wound closure assay.

These results suggest that MMPs do not act as chemoattractants. This reduction in cancer cell invasion was dependent on the dose of antibodies used Figure 4D. Taken together, these in vitro results suggest that astrocyte secreted MMP-2 and MMP-9 proteins partially mediate tumor cell invasion. Intriguingly, each subsequent passage of MDA-MBLuc cells in astrocyte CM increased the invasion competence several fold with passage 5 cells showing an approximate fold increase in invasion compared to passage 1 cells Figure 5A.

We monitored tumor-injected mice using biophotonic imaging at day 0, 2, 4, 6, 8, 10, 12, 14, 21, 28, 35 and 42 of inoculation. Br-Luc cells Figure 5B. By day 14, Br-Luc cells indicating a ten day delay in incidence rate for the brain homing tumor cells Figure 5B and Table S2.

All images of mice are shown in Figure S3. This data collectively suggests that the astrocyte-conditioned tumor cells showed a higher incidence of tumor cells in the brain leading to early mortality. Tumor metastasis was measured by bioluminescent imaging. The decreased normalized photon flux from day 28 was caused by the death of mice Figure S4B and Table S3.

All images of mice are shown in Figure S4A. Together, our data provide evidence that astrocyte secreted MMP-2 and MMP-9 partially mediate astrocyte secretome induced breast cancer metastasis to brain.

Data from our lab provides evidence that the astrocytic secretome influences the local microenvironment, thereby facilitating migration and invasion of breast and lung cancer cells synonymous with metastatic behavior. Because metastasis to the brain is a critical determinant of morbidity, it is vitally important to understand the mechanisms of brain metastasis, thereby facilitating the development of molecular targeted agents that can prevent brain metastasis.

Here, we demonstrate that astrocyte-conditioned tumor cells display highly invasive and metastatic behavior in vitro and in vivo.

MMP-2 and -9 are two factors in the astrocyte secretome that are partially responsible for this response, and blocking MMP-2 and -9 proteins partially prevents the invasion and metastasis of tumor cells in vitro and in vivo. Clinical observations of cancer patients and studies with experimental rodent tumors have concluded that certain tumors produce metastases to specific organs independent of vascular anatomy, and rate of blood flow [25] — [27].

The brain provides a unique environment due to the BBB, a restrictive barrier that is comprised of endothelial cells ECs connected by tight junctions, the basement membrane and astrocytic end-feet processes.

The BBB protects the brain against the entry of most drugs and invasion by microorganisms [29]. In this study, we have identified some of those trophic factors secreted by astrocytes in the soil. In general, astrocytes secrete a wide variety of proteins [32] , [33]. Our filter-cut off studies eliminate most of the cytokines, and heparin agarose binding studies eliminate the remainder of growth factors that have affinity for heparin.

Since filter cut off studies implicate proteins in the 50— kDa range to possess the tumor invasive activity, we anticipate that some of this activity resides in MMPs, more specifically MMP-2 72 and 62 kDa , -9 92 and 82 kDa and -3 54 kDa because the MW of these MMPs fall in the range of 50— kDa.

Indeed, MMP-2 and MMP-9 were detected by gelatin zymography and western blotting in this study, which are consistent with those observed by Gottschall et al where they reported that 1-day-old rat pup brain astrocytes produced MMP-2 after 24 h culture under basal conditions [35].

Also, MMP-2 and -9 have been observed in secretory vesicles in astrocytes [36] implying that some stimulus is responsible for this secretion. Interestingly, astrocytes stimulated with lipopolysaccharide, interleukin-1 alpha or beta, or tumor necrosis factor-alpha for 24 h induce MMP-2 and MMP-9 expression [35] , of which MMP-9 is known to be involved in an MDA-MB brain metastasis model [5].

Strong up-regulation of MMP-9 protein associated with astrocytes was observed in the immediate vicinity of extravasating cancer cells [5] , and is known to promote growth of primary brain tumors by releasing vascular endothelial growth factor VEGF sequestered in the surrounding matrix [37].

Our data here implicates both MMP-2 and -9 secreted by astrocytes in the tumor invasion and metastasis process. Additional activities in the astrocyte CM other than MMPs that contribute to tumor cell invasion are a subject of active investigation in our laboratory.

This hypothesis is further bolstered by the notion that MMPs alone do not contribute much to invasion or migration of tumor cells Figure S2D. What is the mechanism by which MMPs secreted by astrocytes trigger invasion of tumor cells? MMP-2 and -9 secreted by leukemic cells have been shown to increase the permeability of BBB by disrupting tight junction proteins [38]. Similarly, in local ischemia in rat, synthetic MMP inhibitors reverse the MMP-mediated disruption of tight junction proteins in cerebral vessels [39].

These studies imply that secreted MMPs are capable of disrupting BBB thus facilitating invasion into the brain microenvironment. In our study, astrocyte-conditioned tumor cells were injected into the heart. From this site, the tumor cell enters circulation and travels via organs ex: lung en route to brain or directly accesses the brain microenvironment.

Either way, the astrocyte-conditioned tumor cells hone to the brain earlier than non-conditioned tumor cells or cell previously conditioned for brain homing. Because astrocyte-conditioned tumor cells show invasive phenotype, it suggests morphological changes in tumor cells LW, RR unpublished data that influence its ability to invade and metastasize. Alternatively, latent MMP substrates on tumor cells or cells of the tumor microenvironment may be activated upon cleavage by astrocyte secreted MMPs resulting in an invasive phenotype.

Therefore, we favor the morphological alteration model that is dictated in part by genomic changes in the astrocyte-conditioned tumor cells, which is currently under investigation in our laboratory. The outcome of the metastatic process depends on productive interactions of tumor cells with host homeostatic mechanisms [40] , [41].

In the brain, such productive interactions for tumor cells are likely to occur with astrocytes. Although reactive astrocytes surrounding brain metastases has been observed [4] , [8] our data suggests a novel mechanism by which tumor cells when conditioned with astrocyte secretome MMP-2 and -9 becomes highly migratory and invasive leading to tumor cells metastasis to the brain and other sites Figure 7.

We provide evidence for a direct role astrocytes play in altering the immediate tumor microenvironment to facilitate tumor cell invasion leading to increased metastasis to the brain. This study collectively suggests a direct link between astrocytes and tumor progression, and that astrocytes directly influence tumor growth and metastasis. In this model, astrocyte-secreted MMPs and unknown factor s induce cleavage of substrates on tumor cell or surrounding cells in the microenvironment that leads to tumor invasion and metastasis.

TC: tumor cell; SC: surrounding cell. Determine astrocyte-secreted components that mediate tumor cell migration and invasion.

NS: no significance; D S cell invasion in response to astrocyte ultracentrifuge fractionation elutes. Upper panel display invaded cells on the lower surface of the filter; Lower panel is the quantity of the CMs-induced tumor cell invasion.

The resulted medium was submitted to invasion assay 14 h. D MMPs do not possess chemoattractive properties. Biomaterials 81, 72—83, Even-Ram, S. Cell migration in 3D matrix. Curr Opin Cell Biol 17, —, Smalley, K. Animal 42, —, Mosadegh, B. Epidermal growth factor promotes breast cancer cell chemotaxis in CXCL12 gradients. Biotechnol Bioeng , —, Paszek, M. Tensional homeostasis and the malignant phenotype. Cancer Cell 8, —, Campbell, L.

Breast tumor heterogeneity: cancer stem cells or clonal evolution? Cell Cycle 6, — Hughes-Alford, S. Quantitative analysis of gradient sensing: towards building predictive models of chemotaxis in cancer. Curr Opin Cell Biol 24, —, Ichinose, J. Biochem Biophys Res Commun , —, Schulte, A.

Clinical Cancer Research 18, —, Ccr Friedl, P. Tumour-cell invasion and migration: diversity and escape mechanisms. Nat Rev Cancer 3, —, Lammermann, T. Rapid leukocyte migration by integrin-independent flowing and squeezing. Nature , 51—55, Fraley, S. A distinctive role for focal adhesion proteins in three-dimensional cell motility.

Nat Cell Biol 12, —, Gaggioli, C. Fibroblast-led collective invasion of carcinoma cells with differing roles for RhoGTPases in leading and following cells. Nat Cell Biol 9, —, Kalluri, R. Fibroblasts in cancer. Nat Rev Cancer 6, —, Orimo, A.

Loessner, D. Bioengineered 3D platform to explore cell-ECM interactions and drug resistance of epithelial ovarian cancer cells. Kothapalli, C. Theoretical and experimental quantification of the role of diffusive chemogradients on neuritogenesis within three-dimensional collagen scaffolds. Acta Biomater 10, —, Wang, C. A novel in vitro flow system for changing flow direction on endothelial cells.

J Biomech 45, —, Stylianopoulos, T. Diffusion anisotropy in collagen gels and tumors: the effect of fiber network orientation. Biophys J 99, —, Sabeh, F. Protease-dependent versus -independent cancer cell invasion programs: three-dimensional amoeboid movement revisited.

J Cell Biol , 11—19, Guo, Y. Differentiation of clinically benign and malignant breast lesions using diffusion-weighted imaging. Journal of magnetic resonance imaging : JMRI 16, —, Lyng, H. Measurement of cell density and necrotic fraction in human melanoma xenografts by diffusion weighted magnetic resonance imaging. Magnetic resonance in medicine 43, — Download references.

We would like to thank Supriya Nagaraju and Eric Barrientos for their assistance with the time-lapse imaging. Additionally, we would like to thank Prof. Moreover, we acknowledge ASU Graduate and Professional Student Association for funding graduate student travel and research grants for this project.

We would also like to acknowledge the NSF award You can also search for this author in PubMed Google Scholar. All authors reviewed the manuscript. B Distribution of cells was quantified for days 1 and 3. By the third day, there was a shift in cell distribution toward the right indicating a higher cell count further away from the tumor region. C Invasion distance of the tumor front was calculated from the radial distances of the furthest cells from the tumor region.

This work is licensed under a Creative Commons Attribution 4. Reprints and Permissions. Truong, D. Sci Rep 6, Download citation. Received : 13 June Accepted : 07 September Published : 28 September Anyone you share the following link with will be able to read this content:. Sorry, a shareable link is not currently available for this article.

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Download PDF. Subjects Assay systems Breast cancer Cell invasion Lab-on-a-chip. Abstract In this study, to model 3D chemotactic tumor-stroma invasion in vitro , we developed an innovative microfluidic chip allowing side-by-side positioning of 3D hydrogel-based matrices. Introduction Breast cancer is the one of leading causes of cancer-related death among women in the United States 1.

Figure 1. Spatial-organization of ECM and cells. Full size image. Figure 2. Diffusion of molecules through the two regions.

Figure 3. Behavior of breast cancer cells within different ECMs. Figure 5. Breast cancer 3D invasion assay using asymmetric gradients. Figure 6. Figure 7. Figure 8. Figure 9. Analysis of cell morphology. Figure Discussion Despite significant progress, the majority of microfluidic models lack cell and ECM spatial organization that allows for separate manipulation of side-by-side tumor and stromal regions to assess the roles of microenvironmental factors on 3D cancer invasion, in a real-time fashion 25 , 26 , 28 , Conclusions In this work, a new 3D microfluidic platform, designed with separate tumor-stroma entities, was developed to recapitulate 3D cancer cell invasion.

Materials and Methods Microfluidic design and fabrication The microfluidic platform was fabricated using photo- and soft-lithography techniques. Cell culture SUM breast cancer cells was chosen as a suitable cell type to invade through a 3D hydrogel Invasion assay To load the cancer cells into the device for invasion experiments, cells were trypsinized and pelleted into 0.

Statistical analysis All measurements were compiled from three or more independent devices for each experimental condition. Additional Information How to cite this article : Truong, D. References Siegel, R. Article Google Scholar Hanahan, D. Article Google Scholar Knowlton, S. Article Google Scholar Meng, Q. Acknowledgements We would like to thank Supriya Nagaraju and Eric Barrientos for their assistance with the time-lapse imaging. Kamm Authors Danh Truong View author publications.

View author publications. Figure 4. Breast cancer 3D invasion assay. Ethics declarations Competing interests The authors declare no competing financial interests. Electronic supplementary material. Supplementary Information. Supplementary Movie S1. Supplementary Movie S2. Supplementary Movie S3.

Supplementary Movie S4. Supplementary Movie S5.