Interactions of human ovarian tumor cells with human mesothelial cells grown on extracellular matrix

Niedbala, Michael J.; Crickard, Kent; Bernacki, Ralph J.

doi:10.1016/0014-4827(85)90197-1

Cited by 130 publications

(128 citation statements)

References 25 publications

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“…The organs in the peritoneal cavity are normally covered with the peritoneum and subperitoneal connective tissue, whereas the tumor nodules disseminated over the peritoneal surfaces are devoid of such coverings, since the interaction of cancer cells with the peritoneum induces disruption and exfoliation of the mesothelial cells in the early process of peritonal metastasis. 32,33 The peritoneum and underlying fibrous component may act as a barrier against the entry of DNA:PEI complexes through the surface of organs. Interestingly, relatively high luciferase activity was detected within the cells of the peritoneum and underlying fibrous layer (renal capsule), and a significant luciferase activity was observed in the kidney parenchyma without the peritoneal barrier.…”

Section: Discussionmentioning

confidence: 99%

Polyethylenimine-mediated gene transfer into pancreatic tumor dissemination in the murine peritoneal cavity

Aoki¹,

Furuhata²,

Hatanaka³

et al. 2001

Gene Ther

118

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Section: Discussionmentioning

confidence: 99%

Polyethylenimine-mediated gene transfer into pancreatic tumor dissemination in the murine peritoneal cavity

Aoki¹,

Furuhata²,

Hatanaka³

et al. 2001

Gene Ther

118

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“…Previously, several groups [8][9][10][11][12][13][14] had used endpoint assays to show that ovarian cancer cells attach to and invade into mesothelial cell monolayers. This assay is unique in that it uses fluorescently labeled cells to distinguish tumor cells from mesothelial cells, so that the dynamics of these two cell populations can be monitored throughout the assay.…”

Section: Movie 3 Control Ovca433 Spheroids (Red) Invading Into a Mesmentioning

confidence: 99%

“…Previous in vitro experiments demonstrated that primary ovarian cancer cells attach more efficiently to extracellular matrix than to mesothelial cells 8 , and more recent studies showed that primary peritoneal mesothelial cells actually provide a barrier to ovarian cancer cell adhesion and invasion (as compared to adhesion and invasion on substrates that were not covered with mesothelial cells) 9,10 . This would suggest that mesothelial cells act as a barrier against ovarian cancer metastasis.…”

mentioning

confidence: 99%

“…Our protocol was adapted from previously described methods for analyzing ovarian tumor cell interactions with mesothelial monolayers [8][9][10][11][12][13][14][15][16] , and was first described in a report showing that ovarian tumor cells utilize an integrin -dependent activation of myosin and traction force to promote the exclusion of the mesothelial cells from under a tumor spheroid 17 . This model takes advantage of time-lapse fluorescence microscopy to monitor the two cell populations in real time, providing spatial and temporal information on the interaction.…”

mentioning

confidence: 99%

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<em>In vitro</em> Mesothelial Clearance Assay that Models the Early Steps of Ovarian Cancer Metastasis

Davidowitz

Iwanicki

Brugge

2012

JoVE

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Ovarian cancer is the fifth leading cause of cancer related deaths in the United States 1 . Despite a positive initial response to therapies, 70 to 90 percent of women with ovarian cancer develop new metastases, and the recurrence is often fatal 2 . It is, therefore, necessary to understand how secondary metastases arise in order to develop better treatments for intermediate and late stage ovarian cancer. Ovarian cancer metastasis occurs when malignant cells detach from the primary tumor site and disseminate throughout the peritoneal cavity. The disseminated cells can form multicellular clusters, or spheroids, that will either remain unattached, or implant onto organs within the peritoneal cavity 3 (Figure 1, Movie 1).All of the organs within the peritoneal cavity are lined with a single, continuous, layer of mesothelial cells [4][5][6] (Figure 2). However, mesothelial cells are absent from underneath peritoneal tumor masses, as revealed by electron micrograph studies of excised human tumor tissue sections 3,5-7 (Figure 2). This suggests that mesothelial cells are excluded from underneath the tumor mass by an unknown process.Previous in vitro experiments demonstrated that primary ovarian cancer cells attach more efficiently to extracellular matrix than to mesothelial cells 8 , and more recent studies showed that primary peritoneal mesothelial cells actually provide a barrier to ovarian cancer cell adhesion and invasion (as compared to adhesion and invasion on substrates that were not covered with mesothelial cells) 9,10 . This would suggest that mesothelial cells act as a barrier against ovarian cancer metastasis. The cellular and molecular mechanisms by which ovarian cancer cells breach this barrier, and exclude the mesothelium have, until recently, remained unknown.Here we describe the methodology for an in vitro assay that models the interaction between ovarian cancer cell spheroids and mesothelial cells in vivo ( Figure 3, Movie 2). Our protocol was adapted from previously described methods for analyzing ovarian tumor cell interactions with mesothelial monolayers [8][9][10][11][12][13][14][15][16] , and was first described in a report showing that ovarian tumor cells utilize an integrin -dependent activation of myosin and traction force to promote the exclusion of the mesothelial cells from under a tumor spheroid 17 . This model takes advantage of time-lapse fluorescence microscopy to monitor the two cell populations in real time, providing spatial and temporal information on the interaction. The ovarian cancer cells express red fluorescent protein (RFP) while the mesothelial cells express green fluorescent protein (GFP). RFP-expressing ovarian cancer cell spheroids attach to the GFP-expressing mesothelial monolayer. The spheroids spread, invade, and force the mesothelial cells aside creating a hole in the monolayer. This hole is visualized as the negative space (black) in the GFP image. The area of the hole can then be measured to quantitatively analyze differences in clearance activity between cont...

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“…Invasive potential was determined in vitro by transmigration of an ECM (Niedbala et al, 1985). Matrigel invasion chamber plates were purchased from Becton Dickinson/Biocoat (Bedford, MA, USA).…”

Section: Invasion Assaysmentioning

confidence: 99%

STAT3 is required but not sufficient for EGF receptor-mediated migration and invasion of human prostate carcinoma cell lines

Zhou¹,

Grandis

Wells³

2006

Br J Cancer

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Growth factor-induced migration is a rate-limiting step in tumour invasiveness. The molecules that regulate this cellular behaviour would represent novel targets for limiting tumour cell progression. Epidermal growth factor (EGF) receptor (EGFR)-mediated motility, present in both autocrine and paracrine modes in prostate carcinomas, requires de novo transcription to persist over times greater than a few hours. Therefore, we sought to define specific signalling pathways that directly alter cellular transcription. Signal transducer and activator of transcription 3 (STAT3) is activated, as determined by electrophoretic motility shift assays, by EGFR in DU145 and PC3 human prostate carcinoma cells in addition to the motility model NR6 fibroblast cell line. Inhibition of STAT3 activity by antisense or siRNA downregulation or expression of a dominant-negative construct limited cell motility as determined by an in vitro wound healing assay and invasiveness through a extracellular matrix barrier. The expression of constitutively activated STAT3 did not increase the migration, which indicates that STAT3 is necessary but not sufficient for EGFR-mediated migration. These findings suggest that STAT3 signalling may be a new target for limiting prostate tumour cell invasion. In a microarray gene analysis of what transcription units are altered by EGF in a STAT3-dependent manner we found that the expression of motility-limiting VASP protein and the apoptosis nexus caspase 3 were both downregulated upon EGF exposure. These findings suggest a molecular basis for the STAT3 dependence of EGFR-mediated prostate tumour progression.

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Interactions of human ovarian tumor cells with human mesothelial cells grown on extracellular matrix

Cited by 130 publications

References 25 publications

Polyethylenimine-mediated gene transfer into pancreatic tumor dissemination in the murine peritoneal cavity

Polyethylenimine-mediated gene transfer into pancreatic tumor dissemination in the murine peritoneal cavity

<em>In vitro</em> Mesothelial Clearance Assay that Models the Early Steps of Ovarian Cancer Metastasis

STAT3 is required but not sufficient for EGF receptor-mediated migration and invasion of human prostate carcinoma cell lines

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