Next, we evaluated autophagic activity in cells (HepG2, Huh7, and L02) transfected with a small hairpin RNA (shRNA) targeting HMGB1. As shown in Fig. 5A, these cells had markedly diminished Beclin1 protein levels, LC3‐II expression, GFP‐LC3 puncta levels, and autophagosome formation, indicating that HMGB1 is indeed an important factor regulating autophagy in hepatocytes and hepatoma cells. This evidence concerns the gene BECN1 and hepatocellular carcinoma.