MicroRNAs(miRNAs)affect cancer cell glucose metabolism by targeting mRNAs of diverse enzymes that have been implicated in oxidative phosphorylation(OXPHOS)and glycolytic pathways.However,the mechanisms that underlie miRNA-mediated regulation of phosphofructokinase(PFK),a key rate-limiting enzyme in glycolysis,remain largely unknown.Here,we show that miR-128 directly targets PFK liver type(PFKL)in lung cancer cells and regulates endogenous expression of PFKL at both the mRNA and protein levels.In line with this,overexpression of miR-128 decreased glucose uptake and lactate production,as well as increased cellular ATP content.Interestingly,knockdown of miR-128 was shown to promote lung cancer cell growth and colony formation.Moreover,we observed that miR-128 expression inversely correlated with PFKL mRNA levels in clinic lung cancer samples and that increased PFKL expression predicted poor overall survival in lung cancer patients.Mechanistically,we showed that miR-128 regulates PFKL via a feedback loop that involves inhibition of the AKT signaling pathway.Together,our results suggest that miR-128 acts as a metabolic regulator in lung cancer cells that may be therapeutically exploited.