Tracking the Metabolism of 13C L-Aspartic acid in Hepatocellular Carcinoma through NMR Spectroscopy

Hepatocellular carcinoma (HCC) is one of the most common types of liver cancer globally. There is a pressing clinical need for more targeted therapeutics and clear-cut diagnostic tools for early detection and accurate assessment of liver cancer. Cell culture of HCC cells (Huh-7 cell line) was done using DMEM with 10% FBS and doped with 5 mM 13C-aspartic acid for 1hr and 48 hrs. A control without aspartic acid was also prepared and tested. The samples were done in triplicate and measured using a 600 MHz NMR spectrometer. Nuclear Magnetic Resonance (NMR) spectroscopy is a technique that uses the magnetic properties of atomic nuclei to unravel certain molecular compositions. Both cell and media extracts were analyzed to obtain NMR results. NMR results show that aspartic acid entered the tricarboxylic acid (TCA) cycle through oxaloacetate and produced alpha-ketoglutarate, citrate, phosphoglycerate, and a number of other TCA metabolites.