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Comparison of the effects of metformin on MDA-MB-231 breast cancer cells in a monolayer culture and in tumor spheroids as a function of nutrient concentrations
Bizjak, Maruša, 1989- ...
Metabolic pathways of cancer cells depend on the concentrations of nutrients in their micro-environment as well as on the cell-to-cell interactions. Here we examined the effects of glucose, pyruvate ...and glutamine on the sensitivity of MDA-MB-231 cells to metabolic drug metformin using standard 2D culture, in which cells are grown in a monolayer, and 3D tumor spheroids, in which three-dimensional growth of cells better mimics a tumor. To examine effects of nutrients on metformin action, MDA-MB-231 cells were grown in commonly used media (DMEM, MEM and RPMI-1640) that differ mainly in the concentrations of amino acids. We used MTS assay and Hoechst and propidium iodide staining to determine cell number, viability and survival, respectively. We also determined the size of tumor spheroids and assessed effects of nutrients on metformin- stimulated AMP-activated protein kinase activation. Non-essential amino acids suppressed the effects of metformin on MDA-MB-231cells in a 2D culture and in 3D tumor spheroids. Glutamine and pyruvate weakly diminished the effects of metformin in 2D culture. Furthermore, glucose protected tumor spheroids against metformin-induced disintegration. Our results show that nutrient availability must be considered when we evaluate the effects of metformin in 2D culture and in biologically more relevant 3D tumor spheroids.
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