Tracking the effect of BCAT inhibitor on leucine production in glioblastoma cells using carbon-13 NMR spectroscopy

Glioblastoma is an aggressive form of malignant brain tumor in adults, with a 5 year survival rate of only 6.8%. The enzyme branched chain amino acid transaminase (BCAT), responsible for metabolism of branched chain amino acids (BCAA) such as leucine and their ketoacid equivalents e.g. alpha-ketoisocaproic acid (KIC), is found to be upregulated. In this study we have used alpha-KIC as a NMR probe to monitor the effect of BCAT1 inhibitor 2 on alpha-KIC and Leucine metabolism within the SFxL cell line of glioblastoma. BCAT1 inhibitor 2 was determined to have presence within SFxl cells with an IC50 point of around 54 µM via western blotting and XTT assay techniques. Informed by these data we introduced complete media with 70 µM BCAT1 inhibitor 2 concentration to SFxl cells and found through NMR that lessened Leucine production, and increased β-hydroxy β-methyl butyrate production correspond with the earlier BCAT1 inhibitor 2 was introduced to the sample. These results will be discussed with other supporting data