Measurement of Fatty Acid Oxidation Rates in Animal Tissues and Cell Lines

Abstract

While much oncological research has focused on metabolic shifts in glucose and amino acid oxidation, recent evidence suggests that fatty acid oxidation (FAO) may also play an important role in the metabolic reprogramming of cancer cells. Here, we present a simple method for measuring FAO rates using radiolabeled palmitate, common laboratory reagents, and standard supplies. This protocol is broadly applicable for measuring FAO rates in cultured cancer cells as well as in both malignant and nontransformed animal tissues.

NAFLD causes selective CD4+ T lymphocyte loss and promotes hepatocarcinogenesis

“…Fatty acid oxidation was measured according to previous publication 37 . 1- 14 C-linoleic acid and 1- 14 C-palmitic acid were purchased from PerkinElmer.…”

MicroRNA-33–dependent regulation of macrophage metabolism directs immune cell polarization in atherosclerosis

“…C-oleic acid (PerkinElmer) in DMEM, in the presence or absence of etomoxir (25 μM, Sigma-Aldrich), as previously described (67). Oxidation was stopped by the addition of 80 μl of 50% TCA per well, and 14 CO 2 was trapped on Whatman paper disks soaked with 20 μl of 1M NaOH for 1 hour in a fume hood, after which trapped 14 CO 2 was measured by liquid scintillation counting.…”

9‐Oxo‐10(E),12(Z),15(Z)‐Octadecatrienoic Acid Activates Peroxisome Proliferator‐Activated Receptor α in Hepatocytes

“…These experiments were conducted as previously described with some modifications [27][28][29]. In the measurement of 14 C-palmitic acid uptake, the cells were washed with PBS (500 µL/well) and then incubated in assay buffer ( 14 C-palmitic acid, 2 µCi/well; fatty acid free BSA, 2.5 %; and normal palmitic acid, 200 µM) for 3 min at room temperature (1 mL/well).…”