The tumor suppressor p27kip1 get deregulated in aggressive human cancers at the protein level but its precise contribution to the disease remains uncertain. Recently our lab discovered a novel role for p27 in controlling a metabolic switch to amino acids when glucose levels drop. Thus, p27 deregulation provides a growth advantage to cells by allowing them to adapt metabolisms in varied nutrient environments. The switch can be observed by comparing cells with and without p27 when they are subjected to various metabolic inhibitors. Although these results indicate that cells lacking p27 can metabolize amino acids, their identity is not known. To determine which amino acids can support ATP production in p27-/- cells, we varied the identity of amino acids in cell culture media under low glucose levels. Reconstituting cell culture media from base ingredients made it easier to add back specific amino acids individually and in combination. Cells with and without p27 were seeded in 96 well plates under normal conditions, then switched to low glucose and the various amino acids were added. By using CellTiterGlo<sup>TM</sup> assay from Promega, ATP production values can be measured. Results from these studies provide insight into the novel role played by p27 in metabolic switch. By determining which amino acids are being metabolized in the p27-/- cells, it may be possible to identify specific inhibitors to block this process and hence aggressive tumors with deregulated p27 can be targeted.

Metabolism of different amino acids by cells lacking the tumor suppresser p27

2019 Spring National Meeting

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