Interactions of tumor cells with vascular endothelial cell monolayers: a model for metastatic invasion.

Kramer, Randall H.; Nicolson, Garth L.

doi:10.1073/pnas.76.11.5704

Cited by 227 publications

(123 citation statements)

References 30 publications

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“…The prostate epithelial cell then generates a pseudopodial extension, which penetrates the endothelial cell layer, the endothelial cells retract and the prostate cell moves through the endothelial barrier. As with the migration across endothelia observed in melanoma cells, prostate epithelial trans-migration is considerably slower than leukocyte transmigration, with 29% of cells completing the transit within 4 h. This extravasation time is comparable to that observed for melanoma cells (Voura et al, 1998), rat ascites hepatoma cells (Ohigashi et al, 1989) and other tumour cells (Kramer and Nicolson, 1979). However, it has been shown that over 50% of monocytes can cross an endothelium within the first hour of contact (Sandig et al, 1997) without inducing endothelial cell retraction.…”

Section: Discussionsupporting

confidence: 63%

Invasive characteristics of human prostatic epithelial cells: understanding the metastatic process

et al. 2005

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Prostate cancer has a predilection to metastasise to the bone marrow stroma (BMS) by an as yet uncharacterised mechanism. We have defined a series of coculture models of invasion, which simulate the blood/BMS boundary and allow the elucidation of the signalling and mechanics of trans-endothelial migration within the complex bone marrow environment. Confocal microscopy shows that prostate epithelial cells bind specifically to bone marrow endothelial-to-endothelial cell junctions and initiate endothelial cell retraction. Trans-endothelial migration proceeds via an epithelial cell pseudopodial process, with complete epithelial migration occurring after 232743 min. Stromal-derived factor-1 (SDF-1)/CXCR4 signalling induced PC-3 to invade across a basement membrane although the level of invasion was 3.5-fold less than invasion towards BMS (P ¼ 0.0007) or bone marrow endothelial cells (P ¼ 0.004). Maximal SDF-1 signalling of invasion was completely inhibited by 10 mM of the SDF-1 inhibitor T140. However, 10 mM T140 only reduced invasion towards BMS and bone marrow endothelial cells by 59% (P ¼ 0.001) and 29% (P ¼ 0.011), respectively. This study highlights the need to examine the potential roles of signalling molecules and/or inhibitors, not just in single-cell models but in coculture models that mimic the complex environment of the bone marrow.

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Section: Discussionsupporting

confidence: 63%

Invasive characteristics of human prostatic epithelial cells: understanding the metastatic process

et al. 2005

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“…Most works on the transendothelial migration of cancer cells show the formation of gaps in the EC monolayer that allow direct access of TCs to the exposed subendothelial matrix. 21,22,[37][38][39] In this regard, it has been reported that TC-EC interactions induce a rapid EC-EC dissociation, which correlates with a dramatic loss of VE-cadherin staining around the TCs. 34 In contrast, only local changes in VE-cadherin staining at sites of TC-EC contact were reported using a 3-dimensional EC culture system on matrigel, 40 in accordance with our results with collagen gels.…”

Section: Discussionmentioning

confidence: 99%

Regulatory role of tetraspanin CD9 in tumor–endothelial cell interaction during transendothelial invasion of melanoma cells

Longo

Yáñez-Mó

Mittelbrunn

et al. 2001

Blood

108

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Heterotypic interaction among tumor cells (TCs) and endothelial cells (ECs) may play a critical role during the vascular dissemination of neoplastic cells and during pathologic angiogenesis in tumors.To identify molecules involved in these processes, the distribution of vascular junctional proteins was first studied by immunofluorescence at sites of heterologous intercellular contact using TC-EC mosaic monolayers grown on 2-dimensional collagen. Several members of the tetraspanin superfamily, including CD9, CD81, and CD151, were found to localize at the TC-EC contact area. The localization of tetraspanins to the TC-EC heterologous contact area was also observed during the active transmigration of TCs across EC monolayers grown onto 3-dimensional collagen matrices. Dynamic studies by time-lapse immunofluorescence confocal microscopy showed an active redistribution of endothelial CD9 to points of melanoma insertion. Anti-CD9 monoclonal antibodies were found to specifically inhibit the transendothelial migration of melanoma cells; the inhibitory effect was likely caused by a strengthening of CD9-mediated heterotypic interactions of TCs to the EC monolayer. These data support a novel mechanism of tetraspanin-mediated regulation of TC transcellular migration independent of TC motility and growth during metastasis and a role for these molecules in the formation of TC-EC mosaic monolayers during tumor angiogenesis. IntroductionThe dissemination of cells from a primary tumor by invasion and metastasis represents the hallmark of malignance. 1 Invasion is the local dissemination of neoplastic tissue, whereas metastasis is the distant dissemination to secondary places of growth involving the transport of neoplastic cells through the fluid spaces of the body (blood, lymph, cerebrospinal, or peritoneal fluid). 2 Although tumor cell (TC) migration through the extracellular matrix (ECM) is required for invasion, success in metastasis requires migration through the ECM and across host cell layers (transcellular migration). 3 Because most cancer cells reach distant sites by dissemination through blood or lymphatic circulation, transendothelial migration of TCs is a crucial event in vascular metastasis formation. In addition, 2 recent works in human melanoma 4 and colon carcinoma 5 suggesting that tumor vessels are "mosaic" lined by endothelial cells (ECs) and malignant TCs extend the role of TC-EC interactions to the process of tumor angiogenesis. Thus, TC transendothelial invasion and molecules involved in TC-EC interactions could contribute to the formation of new blood vessels in tumors.Tetraspanins comprise a numerous group of proteins that have 4 putative transmembrane domains and that have been implicated in the regulation of cell development, proliferation, activation, and motility. 6,7 It has been suggested that tetraspanins may provide a bridge between ␤1 integrins and other proteins to generate functional complexes that regulate cell-cell and cell-ECM interactions. 8 The expression of many members of the tetraspanin s...

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“…The various components of tumor cell-endothelial cell interaction can be replicated in vitro by challenging a monolayer of human umbilical vein endothelial cells (HUVEC) with cancer cells. Studies performed with electron and phasecontrast microscopy suggest that the in vitro sequence of events fairly represent the in vivo metastatic process 6 . Here, we describe an electricalimpedance based technique that monitors and quantifies in real-time the invasion of endothelial cells by malignant tumor cells.…”

Section: Introductionmentioning

confidence: 99%

A Real-time Electrical Impedance Based Technique to Measure Invasion of Endothelial Cell Monolayer by Cancer Cells

Rahim

Üren

2011

JoVE

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Metastatic dissemination of malignant cells requires degradation of basement membrane, attachment of tumor cells to vascular endothelium, retraction of endothelial junctions and finally invasion and migration of tumor cells through the endothelial layer to enter the bloodstream as a means of transport to distant sites in the host [1][2][3] . Once in the circulatory system, cancer cells adhere to capillary walls and extravasate to the surrounding tissue to form metastatic tumors 4,5 . The various components of tumor cell-endothelial cell interaction can be replicated in vitro by challenging a monolayer of human umbilical vein endothelial cells (HUVEC) with cancer cells. Studies performed with electron and phasecontrast microscopy suggest that the in vitro sequence of events fairly represent the in vivo metastatic process 6 . Here, we describe an electricalimpedance based technique that monitors and quantifies in real-time the invasion of endothelial cells by malignant tumor cells. Giaever and Keese first described a technique for measuring fluctuations in impedance when a population of cells grow on the surface of electrodes 7,8 . The xCELLigence instrument, manufactured by Roche, utilizes a similar technique to measure changes in electrical impedance as cells attach and spread in a culture dish covered with a gold microelectrode array that covers approximately 80% of the area on the bottom of a well. As cells attach and spread on the electrode surface, it leads to an increase in electrical impedance [9][10][11][12] . The impedance is displayed as a dimensionless parameter termed cell-index, which is directly proportional to the total area of tissue-culture well that is covered by cells. Hence, the cell-index can be used to monitor cell adhesion, spreading, morphology and cell density.The invasion assay described in this article is based on changes in electrical impedance at the electrode/cell interphase, as a population of malignant cells invade through a HUVEC monolayer (Figure 1). The disruption of endothelial junctions, retraction of endothelial monolayer and replacement by tumor cells lead to large changes in impedance. These changes directly correlate with the invasive capacity of tumor cells, i.e., invasion by highly aggressive cells lead to large changes in cell impedance and vice versa. This technique provides a two-fold advantage over existing methods of measuring invasion, such as boyden chamber and matrigel assays: 1) the endothelial cell-tumor cell interaction more closely mimics the in vivo process, and 2) the data is obtained in real-time and is more easily quantifiable, as opposed to end-point analysis for other methods. Video LinkThe video component of this article can be found at https://www.jove.com/video/2792/ Protocol 1. Preparation 1. All steps should be performed under sterile conditions in a tissue culture hood. 2. The xCELLigence station is placed in a 37°C incubator in the presence of 5% CO 2 . 3. Use low passage HUVEC cells, preferably no more than passage 6. Also, make sure tha...

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Interactions of tumor cells with vascular endothelial cell monolayers: a model for metastatic invasion.

Cited by 227 publications

References 30 publications

Invasive characteristics of human prostatic epithelial cells: understanding the metastatic process

Invasive characteristics of human prostatic epithelial cells: understanding the metastatic process

Regulatory role of tetraspanin CD9 in tumor–endothelial cell interaction during transendothelial invasion of melanoma cells

A Real-time Electrical Impedance Based Technique to Measure Invasion of Endothelial Cell Monolayer by Cancer Cells

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