β1-integrins regulate the formation and adhesion of ovarian carcinoma multicellular spheroids

Rachael C. Casey, Kathryn M. Burleson, Keith M Skubitz, Stefan E. Pambuccian, Theodore R. Oegema, Laura E. Ruff, Amy P Skubitz

Research output: Contribution to journalArticle

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Abstract

Ovarian carcinoma multicellular spheroids are an in vitro model of micrometastasis whose adhesive abilities have not been elucidated. In this study, we identified adhesion molecules that mediate the formation of ovarian carcinoma spheroids and their subsequent adhesion to extracellular matrix proteins. The NIH: OVCAR5, but not the SKOV3, ovarian carcinoma cell line formed spheroids similar to multicellular aggregates isolated from patient ascitic fluid. NIH:OVCAR5 spheroid formation was augmented by a β1-integrinstimulating monoclonal antibody or exogenous fibronectin, but was inhibited by blocking monoclonal antibodies against the α5- or β1-integrin subunits. By immunohistochemical staining, α2-, α3-, α5-, α6-, and β1-integrin subunits, CD44, and fibronectin were detected in NIH:OVCAR5 spheroids. NIH-OVCAR5 spheroids adhered to fibronectin, laminin, and type IV collagen, and this adhesion was partially inhibited by blocking antibodies against the α5-, α6-, and α2-integrin subunits, respectively. A blocking monoclonal antibody against the β1-integrin subunit completely inhibited adhesion of the spheroids to all three proteins. These results suggest that interactions between the α5β1-integrin and fibronectin mediate the formation of ovarian carcinoma spheroids and that their adhesion to extracellular matrix proteins at sites of secondary tumor growth may be mediated by a complex interaction between multiple integrins and their ligands.

Original languageEnglish (US)
Pages (from-to)2071-2080
Number of pages10
JournalAmerican Journal of Pathology
Volume159
Issue number6
DOIs
StatePublished - Jan 1 2001

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Cellular Spheroids
Integrins
Carcinoma
Fibronectins
Blocking Antibodies
Extracellular Matrix Proteins
Monoclonal Antibodies
Neoplasm Micrometastasis
Collagen Type IV
Ascitic Fluid
Laminin
Adhesives
Staining and Labeling
Ligands
Cell Line
Growth

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β1-integrins regulate the formation and adhesion of ovarian carcinoma multicellular spheroids. / Casey, Rachael C.; Burleson, Kathryn M.; Skubitz, Keith M; Pambuccian, Stefan E.; Oegema, Theodore R.; Ruff, Laura E.; Skubitz, Amy P.

In: American Journal of Pathology, Vol. 159, No. 6, 01.01.2001, p. 2071-2080.

Research output: Contribution to journalArticle

Casey, Rachael C. ; Burleson, Kathryn M. ; Skubitz, Keith M ; Pambuccian, Stefan E. ; Oegema, Theodore R. ; Ruff, Laura E. ; Skubitz, Amy P. / β1-integrins regulate the formation and adhesion of ovarian carcinoma multicellular spheroids. In: American Journal of Pathology. 2001 ; Vol. 159, No. 6. pp. 2071-2080.
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