L-type amino acid transporter 1 (LAT1; SLC7A5) is upregulated in various types of cancer and is often associated with poor prognosis of patients, indicating its pathological significance in disease progression and malignancy. However, our understanding of the mechanisms responsible for such aberrant LAT1 expression in cancer is limited. A previous study reported that LAT1 expression is regulated by hypoxia-inducible factor-2α (HIF-2α) in von-Hippel Lindau (VHL)-deficient renal cell carcinoma cells, which accumulate HIF-2α due to the lack of VHL-dependent proteasomal degradation pathway. It has been thus hypothesized that HIF-2α accumulated under the intratumoral hypoxia may generally contribute to inducing LAT1 expression. In the present study, we assessed this possibility in colorectal cancer (CRC) cells. Experimental hypoxic treatment caused the accumulation of HIF-1α and HIF-2α in CRC cell lines in vitro. Expression of GLUT1 (SLC2A1), a well-known HIFs target, was drastically enhanced in a HIFs-dependent manner. However, LAT1 expression was unresponsive to the accumulation of HIFs under the tested conditions. These results indicate that LAT1 expression is not controlled by HIFs in CRC cells, questioning the expected general roles of the HIFs-mediated hypoxic response in the broad pathological increase of LAT1 in cancer cells.