During tumorigenesis, tumor cells often experience low nutrient conditions. Survival and growth in these conditions can facilitate tumor survival and progression within the body. P27kip1 is a tumor suppressor which is commonly deregulated in aggressive human cancers, and is well known for its role as a cyclin-CDK complex inhibitor. However, our lab has recently identified a novel metabolic role for p27. It was observed that over time, immortalized mouse fibroblast cells containing p27 (p7kip1 +/+) use glucose to depletion, as predicted by the Warburg effect. However, imoratibilzed mouse fibroblasts lacking p27 (p27kip1 -/-) initially used glucose as a carbon source, but then switched to metabolizing media amino acids as glucose levels dropped. This ability to switch carbon sources may facilitate tumor progression under glucose limiting conditions, such as those found in early tumorigenesis. The effect of glucose levels on this metabolic switch has been previously characterized, but the effect of amino acids levels on the cell’s metabolism has not been studied. To examine the effect of amino acid concentrations on the metabolism of p27+/+ and p27-/- cells, fibroblasts were distributed in a 96 well plate and cultured under normal conditions overnight to allow cells to attach. The following day cells were treated with media with varying glucose and amino acid levels for different timepoints. Metabolic inhibitors will also used to confirm which metabolic pathways were in use. To analyze the effect of the culture conditions, ATP levels were measured using a luciferase-based light emission assay from Promega. Examining these cells responses to different amino acid levels will help to further characterize the metabolic role of p27kip1, which may add to our understanding of the survival and progression of tumor cells.

Effect of amino acid concentration on metabolism in cells lacking P27kip1

2019 Spring National Meeting

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