We previously demonstrated that human hepatocellular carcinoma‐derived HuH‐7 cells stimulated with interleukin‐1β (IL‐1β) produce α1‐acid glycoprotein (AGP) with increased amounts of sialyl Lewis X (sLeX) antigen, although the mechanism remained obscure. Here, we report our investigation of the mechanism. sLeX expression on HuH‐7 cells was induced 2.5 times more after 48 h stimulation with 100 U/mL IL‐1β compared with control, as indicated by anti‐sLeX antibody binding. Furthermore, expression of 2,3‐sialylated N‐acetyllactosamine increased gradually up to 48 h after IL‐1β stimulation; this preceded the increase in sLeX expression. Increases in α2,3‐sialyltransferase activity also preceded increases in α1,3‐fucosyltransferase activity. Furthermore, mRNA levels of ST3Gal IV, FUT IV and VI in HuH‐7 cells stimulated with IL‐1β were increased at 2–4 h, while increases in FUT VI mRNA level occurred gradually after 24 h. IL‐1β‐induced sLeX expression on HuH‐7 cells was suppressed by transfection of gene‐specific small interference RNAs against FUT VI and ST3Gal IV but not against FUT IV and ST3Gal III. These data results that IL‐1β induces expression of sLeX on HuH‐7 cells by enhanced expression of FUT VI and ST3Gal IV gene.