Our research centered around acute myelogenous leukemia (AML) and the Warburg Effect. The Warburg Effect describes the tendency of proliferating cells and cancer cells to convert glucose into lactate despite adequate levels of oxygen. As it has been previously revealed that leukemia cells produce lactate even in the presence of oxygen, we hypothesized that AML cells specifically are dependent on aerobic glycolysis and that treatment with dichloroacetate will stimulate apoptosis. We further hypothesized that apoptosis would be the result of the cells switching from aerobic glycolysis to oxidative phosphorylation, resulting in an increase in mitochondrial activity and reactive oxygen species and a decrease in survival proteins Bcl-2 and survivin.
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Several cytotoxicity assays were performed in order to determine the toxicity of dichloroacetate. Using 24 well culture plates, an equal number of cells were treated with 0, 10, 20, 30, 40, 50, and 60 mM concentrations of dichloroacetate in triplicate. Viable cell counts were performed after 24, 48, and 72 hours of treatment using a hemocytometer and trypan blue exclusion dye. The final results were reported as the mean plus the standard error of the mean of all trials. In order to detect any changes in Bcl-2 and survivin levels due to treatment, western blotting was conducted using cells from each dichloroacetate concentration after 48 hours of treatment. Blots were stained with Ponceau S before probing with antibody to control for protein loading. To detect mitochondrial activity, hydrogen peroxide concentrations were to be measured using an Amplex Red kit. Each treatment would be measured after 24 and 48
Several cytotoxicity assays were performed in order to determine the toxicity of dichloroacetate. Using 24 well culture plates, an equal number of cells were treated with 0, 10, 20, 30, 40, 50, and 60 mM concentrations of dichloroacetate in triplicate. Viable cell counts were performed after 24, 48, and 72 hours of treatment using a hemocytometer and trypan blue exclusion dye. The final results were reported as the mean plus the standard error of the mean of all trials. In order to detect any changes in Bcl-2 and survivin levels due to treatment, western blotting was conducted using cells from each dichloroacetate concentration after 48 hours of treatment. Blots were stained with Ponceau S before probing with antibody to control for protein loading. To detect mitochondrial activity, hydrogen peroxide concentrations were to be measured using an Amplex Red kit. Each treatment would be measured after 24 and 48