Application Note 34

Fluxing Through Cancer: Tracking the Fate of ¹³C-Labeled Energy Sources Glucose and Glutamine in Cancer Cells and Mouse Tumors

Costas A. Lyssiotis,^1,2 Susanne B. Breitkopf,^1,2 Min Yuan,¹ Gary Bellinger,¹ John M. Asara^1,2

1. Beth Israel Deaconess Medical Center, Boston, MA 02215 USA
2. Harvard Medical School, Boston, MA 02215 USA

Abstract

Glucose and glutamine provide the primary energy sources for cell growth and proliferation. To study metabolic reprogramming, we used D-glucose (¹³C₆, 99%) (CLM-1396) and L-glutamine (¹³C₅, 99%) (CLM-1822-H) to target and track the diversion of these molecules into several metabolic pathways, including glycolysis, the TCA cycle, the pentose phosphate pathway, the metabolism of amino acids and nucleotides, etc. in both cell lines and mouse tumors. We use a positive/negative ion polarity switching single-column SRM experiment during a 15-minute acquisition. For in vivo labeling experiments, D-glucose (¹³C₆, 99%) or L-glutamine (¹³C₅, 99%) solutions were delivered to tumors via intraperitoneal injection (IP) or jugular delivery and compared. Metabolites were extracted from cells or tumor tissues using 80% methanol. Metabolomics were performed on a AB/SCIEX 5500 QTRAP in SRM mode using amide XBridge HILIC chromatography with Q1/Q3 transitions for both the unlabeled and ¹³C-labeled metabolites with separate methods for glucose and glutamine.

Read more by downloading the application note.