Previously, we reported the expression and function of system L amino acid transporter in KB human oral epidermoid carcinoma cells. In the present study, therefore, we investigated the expression and function of system L amino acid transporter in human normal oral keratinocytes (HNOK) and compared the expressions and functions of system L amino acid transporters in HNOK and KB cells. The HNOK expressed L-type amino acid transporter 1 (LAT1) and L-type amino acid transporter 2 (LAT2) with their subunit 4F2hc in the plasma membrane but the expression of LAT1 was very weak, which is in contrast to the KB cells expressing LAT1 but not LAT2 with the 4F2hc in the plasma membrane. The [14C] L-leucine uptake by HNOK, as well as KB cells, was inhibited by the system L selective inhibitor BCH. The majority of [14C] L-leucine uptake was, therefore, mainly mediated by LAT2 in the HNOK and by LAT1 in the KB cells. These results suggest that the transport of neutral amino acids including several essential amino acids into the HNOK and KB cells are mainly mediated by LAT2 and LAT1, respectively. The specific inhibition of LAT1 in oral cancer cells could be a new rationale for anti-cancer therapy.