The epidermal growth factor is a regulator of a wide variety of processes in various cell systems. Hepatocytes are important sites in the body's metabolism and function. Glucose transporter-2 (GLUT-2) is a major transporter that is expressed strongly in hepatocytes. Therefore, this study examined the effect of EGF on GLUT-2 and its related signal cascades in primary cultured chicken hepatocytes. EGF decreased the [³H]-deoxyglucose uptake in a dose- and time-dependent manner (> 10 ng/ml, 2 hr). AG 1478 (an EGF receptor antagonist), genistein and herbimycin A (tyrosine kinase inhibitors) blocked the EGF-induced decrease in the [³H]-deoxyglucose uptake, which correlates with the GLUT-2 expression level. In addition, the EGF-induced decrease in the GLUT-2 protein expression was inhibited by staurosporine, H-7, or bisindolylmaleimide I (protein kinase C inhibitors), PD 98059 (a MEK inhibitor), SB 203580 (a p38 MAPK inhibitor), and SP 600125 (a JNK inhibitor), suggesting a role of both PKC and MAPKs (p44/42 MAPK, p38 MAPK and JNK). In particular, EGF increased the translocation of PKC isoforms (, 1, , , and ) from the cytosol to the membrane fraction and increased the activation of p44/42 MAPK, p38 MAPK and JNK. Moreover, the PKC inhibitors blocked the EGF-induced phosphorylation of three MAPKs. In conclusion, EGF decreases the GLUT-2 expression level via the PKC-MAPKs signal cascade in chicken hepatocytes.