ABSTRACTS – ORAL PRESENTATIONS - AMCA, spol. s r.o.

of molecular biology we detected changes of proliferation (CyQUANT), differentiation
(alkaline phosphatase activity) and cell death (phosphatidylserine externalisation,
caspase activation, mitochondrial membrane potential, PARP cleavage). These changes
were accompanied by altered expression of some apoptotic (caspase 9) and autophagic
(LC3, p62) proteins. Application of fatty acids also affected lipid metabolism regulating
proteins (fatty acid synthase, caveolin-1) as well as cell ability to incorporate and store
fatty acids (FAT/CD36, lipid droplet accumulation).
In summary, our data suggest participation of PPARγ and specific signaling molecules
in the effects of NaBt, DHA and their combination on colon cancer epithelial cell
differentiation and death.
Acknowledgements
This work was supported by grants Nos. P301/11/1730, 13-097665 of the Czech Science
Foundation, NT 11205-5/2010 and MSM0021622430 Ministry of Education, Youth and
Sports.
P37. INHIBITORS OF CELLULAR ENERGY METABOLISM CAN ACTIVATE PRO-SURVIVAL
SIGNALING IN MALIGNANT MELANOMA CELLS
Amandine Verlande, Stjepan Uldrijan
Department of Biology, Faculty of Medicine, Masaryk University Brno, Czech Republic
Metastatic melanoma is a type of cancer that is notoriously difficult to treat, with only
a minority of patients responding to standard chemotherapeutics. In a search for new
drugs with anti-melanoma activity we investigated the effect of inhibitors of cellular
energy metabolism on malignant melanoma cell survival and the activity of several
important intracellular signaling pathways.
We used three different inhibitors of mitochondrial oxidative phosphorylation (OXPHOS):
Rotenone, CCCP and Oligomycin A, in combinations with two hexokinase inhibitors
(glycolysis pathway): 2-deoxy-D-glucose (2DG) and 5-thio-D-glucose (5TG) in both BRAFand
NRAS-mutated melanoma cells.
Our results show that treatment with the OXPHOS inhibitors leads to cell death that
can be prevented when 2DG or 5TG is added at low, non-toxic concentrations. We show
that these drug combinations induce activation of AKT and AMPK pro-survival pathways.
Even though AKT is strongly activated by the combinations, the mTORC1 complex seems
to be inactive as we observed a simultaneous upregulation of autophagy and inhibition
of protein synthesis.
Taken together, these unexpected results suggest that the combination of OXPHOS and
hexokinase inhibitors can activate pro-survival pathways in malignant melanoma and
prevent cancer cell death.
Analytical Cytometry VII 131