Epithelial-mesenchymal transition in the model of breast carcinoma stem cells in vitro.

Project leader: Ján Sedlák
Project duration: 2006 - 2009

The goal of the project is a characterization of signaling pathways modulation involved in EMT using the in vitro model of breast cancer stem cell (CSC) during different cultivation conditions. The breast CSC will be isolated from cell lines and cultivated as mammospheres. For EMT induction the CSC will be cultivated adherent on plastic (2D cultures) and in the collagen gel (3D cultures) form in the presence of EMT inducers. In parallel, cultivation in hypoxic thermostat allows to compare the marker expression in both conditions. Specific monoclonal antibodies against the phosphorylated form of signaling proteins, as well as treatment with specific pharmacological inhibitors of signaling pathways will be used. We will follow the doubling time kinetics, viability, modulation of selected specific epithelial and mesenchymal markers, mitochondrial potential changes, redox status, and intracellular pH. These results will be compared with retrospective study results, using paraffin-embedded breast carcinoma samples, characterized with the aid of laser scanning cytometer (EMT markers, cell cycle, apoptosis, hypoxia). Markers of stemness and EMT will be used for the detailed tumor characteristics and correlated with clinico-pathological data and the treatment outcome.

Publications