Medizinische Universität Graz Austria/Österreich - Forschungsportal - Medical University of Graz
Gewählte Publikation:
Fuertinger, S; Niesner B.
Die Etablierung von MCF-7 Zelllinien mit Genom-integrierter Expression von GIRK1 und GIRK4
[ Diplomarbeit ] Medical University of Graz; 2011. pp. 50
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- Autor*innen der Med Uni Graz:
- Betreuer*innen:
- Bauernhofer Thomas
- Schreibmayer Wolfgang
- Altmetrics:
- Abstract:
- G protein activated inwardly rectifying potassium channels (GIRKs) represent a connection between membrane potentials and the presence of extra cellular signal molecules. They play an important role in the humoral regulation of cardiac activity, pancreatic insulin distribution and the neurotransmitter regulated neural activity. As noted in recent years, GIRKs have a significant influence on the pathophysiology of breast cancer, particularly proliferation and metastasis. A number of GIRK subtypes have already been detected and identified in cultured breast cancer cells, versus benign breast epithelial cells. The influence of GIRK1 and GIRK4 on morphology and vital signs of breast cancer cells was examined in the breast cancer cell line MCF-7. Transfection of GIRKs is the most basic step for such attempts, the aim of this work was to establish new cell lines which express the GIRK1, or GIRK4 protein stably. The selection of stable transfectants was carried out using the antibiotic G-418 (Geneticin). GIRK proteins with the yellow form of green fluorescence protein (eYFP) were coupled, transfection success was verified using confocal microscopy. Sorting of stable transfected cells was carried out usisng FACS analysis (fluorescence activated cell sorting), which spawned seven GIRK1 clones (A-G) and five GIRK4 clones (2-6). These were then morphologically characterized. In the context of transient transfection subcellular distribution of fluorescence, which was located in GIRK1 in the intracellular area and on GIRK4 in the cell membrane, seemed to be characteristic for each protein. Confocal microscopic investigation of the clones achieved however, that the expression behavior and the growth pattern of cells with strong expression has changed. Further experiments on vital signs of this cell lines are planned.