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Molecular Medicine

, Volume 13, Issue 1–2, pp 79–88 | Cite as

Phosphorylation of FAK, PI-3K, and Impaired Actin Organization in CK-positive Micrometastatic Breast Cancer Cells

  • Galatea Kallergi
  • Dimitris Mavroudis
  • Vassilis Georgoulias
  • Christos Stournaras
Research Article

Abstract

Several markers have been used to detect circulating tumor cells (CTC) in the peripheral blood of patients with breast cancer. However, analysis of activated signaling kinases in CTC implicated in cellular transformation, migration, and survival has not been addressed so far. In the present study, we focused on the phenotypic profile of micrometastatic cells in peripheral blood mononuclear cells (PBMC) preparations from 45 breast cancer patients. PBMC cytospins from 28 cytokeratin (CK)-positive and 17 CK-negative samples were assessed for the expression of phosphorylated FAK (p-FAK), phosphorylated PI-3 kinase (p-PI-3K), and HER2 using confocal laser scanning microscopy. The expression of p-FAK was documented in all 28 CK-positive samples, while all 17 CK-negative samples were tested negative for p-FAK. Immunomagnetic separation using EpCAM antibody fully confirmed these findings, implying a sound correlation for the co-expression of the two molecules. Interestingly, 15 of 28 CK- and p-FAK-positive samples also expressed the HER2 oncoprotein. p-PI-3K was documented in 15 of 17 CK- and p-FAK-positive samples. Immunoblot analysis of micrometastatic cells in co-culture with PBMC confirmed the specific expression of both p-FAK and p-PI-3K. Finally, impaired actin organization was apparent in CK- and p-FAK/p-PI-3K-positive samples, comparable to that observed in MCF-7 human breast cancer cells. Our findings provide strong evidence that micrometastatic cells express activated signaling kinases, which may regulate migration mechanisms, supporting the presumption of their malignant and metastatic nature.

Notes

Acknowledgments

The European Social Fund and National recourses supported this work.

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Copyright information

© Feinstein Institute for Medical Research 2007

Authors and Affiliations

  • Galatea Kallergi
    • 1
  • Dimitris Mavroudis
    • 2
  • Vassilis Georgoulias
    • 2
  • Christos Stournaras
    • 1
  1. 1.Department of BiochemistryUniversity of Crete Medical SchoolHeraklion VoutesGreece
  2. 2.Department of Medical OncologyUniversity Hospital of HeraklionHeraklionGreece

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