Establishment of an in vitro assay to measure the invasion of ovarian carcinoma cells through mesothelial cell monolayers

Rachael C. Casey, Kimberly A. Koch, Theodore R. Oegema, Keith M Skubitz, Stefan E. Pambuccian, Suzanne M. Grindle, Amy P Skubitz

Research output: Contribution to journalArticle

24 Citations (Scopus)

Abstract

Ovarian carcinoma is the leading cause of gynecological cancer deaths in the United States. Secondary tumor growths form by tumor cell invasion through the mesothelial lining of the peritoneal cavity and peritoneal organs. To study this interaction, we developed a dye-based in vitro model system in which mesothelial cells were grown as confluent monolayers, permeabilized, and then co-cultured with ovarian carcinoma cells for up to seven days. The mesothelial cells were then stained with trypan blue dye, which enabled the visualization of ovarian carcinoma cell invasion through the monolayers of mesothelial cells. Ovarian carcinoma cell invasion was inhibited for up to 7 days by the addition of GRGDSP peptides, a blocking monoclonal antibody against the β1 integrin subunit, or blocking monoclonal antibodies against matrix metalloproteinases 2 and 9. Cell invasion was also inhibited by hyaluronan and GM6001, a chemical inhibitor of matrix metalloproteinases. Differential gene expression of matrix metalloproteinases, tissue inhibitors of matrix metalloproteinases, and disintegrins were observed in primary ovarian carcinoma tumors and secondary metastases, compared to normal ovaries. Taken together, these results suggest that complex interactions between integrins, disintegrins, matrix metalloproteinases, and tissue inhibitors of matrix metalloproteinases may mediate ovarian carcinoma cell invasion, and that the dye-based assay described herein is a suitable model system for its study.

Original languageEnglish (US)
Pages (from-to)343-356
Number of pages14
JournalClinical and Experimental Metastasis
Volume20
Issue number4
DOIs
StatePublished - Jul 2 2003

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Carcinoma
Matrix Metalloproteinase Inhibitors
Disintegrins
Tissue Inhibitor of Metalloproteinases
Blocking Antibodies
Coloring Agents
Matrix Metalloproteinases
Integrins
glycyl-arginyl-glycyl-aspartyl-seryl-proline
Neoplasms
Monoclonal Antibodies
In Vitro Techniques
Trypan Blue
Matrix Metalloproteinase 2
Matrix Metalloproteinase 9
Peritoneal Cavity
Hyaluronic Acid
Ovary
Neoplasm Metastasis
Gene Expression

Keywords

  • Cell invasion
  • Integrins
  • Mesothelial cells
  • Ovarian carcinoma

Cite this

Establishment of an in vitro assay to measure the invasion of ovarian carcinoma cells through mesothelial cell monolayers. / Casey, Rachael C.; Koch, Kimberly A.; Oegema, Theodore R.; Skubitz, Keith M; Pambuccian, Stefan E.; Grindle, Suzanne M.; Skubitz, Amy P.

In: Clinical and Experimental Metastasis, Vol. 20, No. 4, 02.07.2003, p. 343-356.

Research output: Contribution to journalArticle

Casey, Rachael C. ; Koch, Kimberly A. ; Oegema, Theodore R. ; Skubitz, Keith M ; Pambuccian, Stefan E. ; Grindle, Suzanne M. ; Skubitz, Amy P. / Establishment of an in vitro assay to measure the invasion of ovarian carcinoma cells through mesothelial cell monolayers. In: Clinical and Experimental Metastasis. 2003 ; Vol. 20, No. 4. pp. 343-356.
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