To understand the role of pyruvate metabolism in the metabolic adaptation of HCC cells under glutamine depletion, we performed stable‐isotope tracing studies with uniformly labelled [U‐13C3] sodium pyruvate in PDHA, PC and PDHB KO clones by gas chromatography‐mass spectrometry (GC‐MS) analysis. This evidence concerns the gene PDHB and hepatocellular carcinoma.