Glycolysis maintains AMPK activation in sorafenib-induced Warburg effect

Hepatocellular carcinoma (HCC) is the second deadly cancer in the world and still lacks curative treatment. Aerobic glycolysis, or Warburg effect, is a major resistance mechanism induced by first-line treatment of HCC, sorafenib, and is regulated by the master regulator of metabolism, AMPK. Activation of AMPK is required for resistance; however, activation dynamics of AMPK and its regulation is rarely studied. Engineering cells to express an AMPK activity biosensor, we monitor AMPK activation in single HCC cells in a high throughput manner during sorafenib-induced drug resistance. Sorafenib induces transient activation of AMPK, duration of which is dependent on glucose. Inhibiting glycolysis shortens AMPK activation; whereas increasing glycolysis increases its activation duration. Our data highlight that activation duration of AMPK is important for cancer evasion of therapeutic treatment and glycolysis is a key regulator of activation duration of AMPK. •AMPK activity is monitored in single cancer cells in a high throughput manner during sorafenib-induced drug resistance.•Sorafenib induces transient activation of AMPK, duration of which is dependent on glucose.•Inhibiting glycolysis shortens AMPK activation, whereas increasing glycolysis increases its activation duration.•Prolonged activation of AMPK is important for evasion of sorafenib-induced cell killing in hepatocellular carcinoma.