Long-noncoding RNA (lncRNA), particularly LBX2-AS1, plays important roles in the occur-rence and development of cancer. Aerobic glycolysis is important in the uncontrolled growth of tumor cells. However, the molecular mechanisms of its involvement remain controversial and have not been widely studied in esophageal cancer. Here, we examined the effects of knockdown of lncRNA LBX2-AS1 on the level of aerobic glycolysis and esophageal cancer progression. The esophageal cancer cell lines KYSE-150 and KYSE170, along with si-LBX2-AS1, which shows stable expression and a high knockdown efficiency, were used to ex-amine the effects of knocking down lncRNA LBX2-AS1 on the activity, proliferation, metastasis, and aerobic glycolysis levels of cells. The LBX2-AS1 signaling pathway involved in promoting esophageal cancer was evaluated using quantitative reverse transcription-PCR and western blotting. The results demonstrated that knockdown of lncRNA LBX2-AS1 inhibited the proliferation, metastasis, and glycolytic effects of esophageal squamous carcinoma cells, as confirmed based on the expression of a key metastatic protein (N-cadherin) and key glycolytic enzymes (hypoxia-inducible factor-1a and pyruvate kinase M2). Knockdown of LBX2-AS1 pro-moted the inhibitory effect of miR-491-5p on pyruvate kinase M2, thereby inhibiting the growth of esophageal cancer cells. In summary, knockdown of lncRNA LBX2-AS1 can target miR-491-5p/pyruvate kinase M2 to reduce glycolysis levels and inhibit the development of esophageal cancer. This lncRNA shows potential as a targeted agent for treating esophageal cancer.