To further test the hypothesis, we performed the corresponding function experiments in ESCC cells, we found that TMEM161B‐AS1 overexpression suppressed cell proliferation, invasion and glycolysis, coupled with increased HIF1AN expression as well as reduced expressions of glycolysis‐related proteins HIF‐1α, HK2, PFKM and LDHA, and converse data were obtained when TMEM161B‐AS1 was depleted. The gene discussed is PFKM; the disease is esophageal squamous cell carcinoma.