Hypoxia is a common characteristic of solid tumors. Previous studies have reported that the tumor invasion-associated factor, AMPK-related protein kinase 5 (ARK5), is associated with a poor prognosis in colon cancer. However, whether or not ARK5 is involved in hypoxia is unclear. The aim of present study was to investigate the association between the expression of ARK5 and that of hypoxia-inducible factor 1-α (HIF1-α). Samples from 60 patients with colon cancer were collected and immunohistochemistry was used to detect the expression of ARK5 and HIF1-α within them. Western blot analysis and reverse transcription polymerase chain reaction were used to detect the expression of ARK5 in an SW480 cell line under hypoxic conditions. Cell Counting kit-8 and Transwell assays were used to study the function of ARK5 under hypoxic conditions. According to the immunohistochemistry results, ARK5 and HIF1-α staining was significantly associated with Tumor-Node-Metastasis stage, tumor grade, lymph node metastasis and liver metastasis. Spearman's correlation analysis revealed a correlation between the expression of ARK5 and that of HIF1-α. This finding was also verified under hypoxic conditions in the SW480 cell line, in which the expression of ARK5 increased over time. Further cellular function experiments revealed that suppression of ARK5 inhibited cell viability and migration under hypoxic conditions. The present study has suggested that ARK5 expression in colon cancer cells is upregulated by HIF1-α under hypoxic conditions and that ARK5 serves an important role in cell proliferation and migration under hypoxic stress.