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Mesenchymal-to-epithelial transition determinants as characteristics of ovarian carcinoma effusions

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Abstract

The present study investigated the intracellular regulation of E-cadherin in ovarian carcinoma. E-cadherin expression and regulation by Snail and Pak1 were studied in ES-2 and OVCAR-3 ovarian cancer cells in vitro. Twist1, Zeb1 and Vimentin mRNA expression and HIF-1α protein expression were analyzed in 80 and 189 clinical specimens, respectively. OVCAR-3 cells incubated with an anti-E-cadherin antibody formed smaller and looser spheroids compared to controls. Snail silencing using Small Hairpin RNA in ES-2 cells reduced invasion and MMP-2 activity, with unaltered cellular morphology. Using dominant negative (DN) and constitutively active (CA) Pak1 constructs, we found that DN Pak1 ES-2 and OVCAR-3 clones had reduced attachment to matrix proteins, invasion and MMP-2 activity compared to CA and wild-type cells. DN Pak1 ES-2 cells also bound less to LP9 mesothelial cells. DN Pak1 OVCAR-3 cells had lower Vimentin levels. Snail expression was lower in cultured effusions compared to primary carcinomas, and was cytoplasmic rather than nuclear. Twist1 (P < 0.001), Zeb1 (P = 0.003) and Vimentin (P = 0.03) mRNA expression was significantly higher in solid metastases compared to primary carcinomas and effusions. HIF-1α protein expression was lower in effusions compared to primary carcinomas and solid metastases (P = 0.033). Our data suggest that the previously reported E-cadherin re-expression in ovarian carcinoma effusions is regulated by Pak1. The transient nature of E-cadherin expression during ovarian carcinoma progression is probably the result of partial epithelial-to-mesenchymal transition (EMT) and the reverse process of mesenchymal-to-epithelial-like transition (MET). Expression of the EMT-related molecules Twist, Zeb1, Vimentin and HIF-1α is anatomic site-dependent in ovarian carcinoma.

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Abbreviations

EMT:

Epithelial-to-mesenchymal transition

MET:

Mesenchymal-to-epithelial transition

OC:

Ovarian carcinoma

DA:

Dominant negative

CA:

Constitutively active

MMP:

Matrix metalloproteinases

TMA:

Tissue microarrays

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Acknowledgments

This work was supported by the Inger and Jon Fredriksen Foundation for Ovarian Cancer Research.

Reuven Reich is affiliated with the David R. Bloom Center for Pharmacy and the Brettler Center for Pharmacology at the Hebrew University of Jerusalem.

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Authors and Affiliations

  1. Institute of Drug Research, School of Pharmacy, Faculty of Medicine, The Hebrew University of Jerusalem, 91120, Jerusalem, Israel

    Sivan Elloul, Olga Vaksman & Reuven Reich

  2. Department of Pathology, Norwegian Radium Hospital, Oslo University Hospital, Montebello, 0310, Oslo, Norway

    Helene Tuft Stavnes & Ben Davidson

  3. Gynecology and Obstetrics, Norwegian Radium Hospital, Oslo University Hospital, 0310, Oslo, Norway

    Claes G. Trope

  4. The Medical Faculty, University of Oslo, 0316, Oslo, Norway

    Claes G. Trope & Ben Davidson

Authors
  1. Sivan Elloul
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  2. Olga Vaksman
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  3. Helene Tuft Stavnes
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  4. Claes G. Trope
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  5. Ben Davidson
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  6. Reuven Reich
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Correspondence to Ben Davidson or Reuven Reich.

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Elloul, S., Vaksman, O., Stavnes, H.T. et al. Mesenchymal-to-epithelial transition determinants as characteristics of ovarian carcinoma effusions. Clin Exp Metastasis 27, 161–172 (2010). https://doi.org/10.1007/s10585-010-9315-2

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  • DOI: https://doi.org/10.1007/s10585-010-9315-2

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