Hypoxia‑inducible‑factor 1α (HIF‑1α) is a marker for poor prognosis in the majority of the cancer types, and it has been revealed to be essential for maintaining cancer stem cells (CSCs). In the present study, it was determined that the expression of HIF‑1α and CSC‑related genes under hypoxic conditions was upregulated. Stable knockdown of HIF‑1α significantly inhibited cell proliferation, migration and tumour growth in vivo in oesophageal squamous cell carcinoma (ESCC). A previous study revealed that the Wnt/β‑catenin pathway may play a key role in maintenance and progression of CSCs. Therefore, it was also revealed that stable knockdown of HIF‑1α reduced the formation of spheroid body cells, the expression of CSC‑related genes and Wnt/β‑catenin pathway‑related target genes, as well as the activity of the Wnt/β‑catenin pathway. Collectively, the present results indicated that HIF‑1α may regulate the stemness of ESCC by activating the Wnt/β‑catenin pathway.