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- W1454298055 abstract "4473 De novo fatty acid synthesis is critical for tumor cell survival, and small molecule inhibitors of this pathway may be useful anti-cancer agents. In this report we sought to evaluate the contributions of two pathway enzymes, acetyl-CoA carboxylase 1 (ACC1) and fatty acid synthase (FAS) on the survival of HCT-116 colon cancer cells. To this end, we performed a kinetic analysis of FAS and ACC1 inhibition using siRNA transfections with two oligonucleotide sequences per target at various concentrations (0, 30 and 60 nM). Cell death (by apoptosis) and viability were assessed and plotted as a function of lipogenesis under the various transfection conditions in order to obtain a specific kinetic profile for knockdown of each enzyme. The resulting curves allowed for a direct comparison of FAS and ACC1 in terms of their significance in maintaining tumor cell viability. A gradual decrease of lipid biosynthesis by as much as 50% was achieved with siRNA-mediated knockdown of either FAS or ACC1 in HCT-116 cells. Reduced lipid biosynthesis correlated well with the expression of either FAS or ACC1. Furthermore, the changes in apoptosis and viability induced by the siRNA titrations, when plotted against lipogenesis, revealed an identical kinetic profile for both FAS and ACC1. For example, following a reduction in lipid biosynthesis by 25%, about 20% of cells stained positive for apoptosis compared to about 3% in control cells, irrespective of which target was knocked down. These findings strongly support the concept that a decrease in de novo lipogenesis, rather than perturbations in pathway intermediates, causes apoptosis in tumor cells when the FAS pathway is inhibited. It is generally difficult to compare targets in a pathway by using single-point siRNA transfections. One reason for this is that knockdown efficiency, and therefore the resulting functional effects, are likely to vary between different targets. Here, we have applied a kinetic approach using siRNA titrations, which allowed us to directly compare enzymes in the FAS pathway with respect to their role in controlling lipogenesis and tumor cell viability. Our results show that FAS and ACC1 are equally important enzymes for controlling lipogenesis and viability in cultured HCT-116 cancer cells." @default.
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- W1454298055 date "2007-05-01" @default.
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- W1454298055 title "Kinetic analysis of the fatty acid synthesis pathway in HCT-116 colon cancer cells: role of FAS and ACC1 in maintaining tumor cell viability and proliferation" @default.
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