The influence of the enhancer of split- and hairy-related protein-2 (SHARP-2) transcriptional repressor on the expression of rat phosphoenolpyruvate carboxykinase (PEPCK) gene was examined. When H4IIE cells were treated with epigallocatechin gallate, a green tea constituent, an increase in SHARP-2 mRNA levels and a decrease in PEPCK mRNA levels were observed. The adenovirus-mediated overexpression of SHARP-2 in H4IIE cells and primary cultured rat hepatocytes led to a decrease in the levels of PEPCK mRNA. Finally, when a SHARP-2 expression plasmid was transiently transfected with various reporter plasmids into MH1C1 cells, the promoter activity of a PEPCK reporter plasmid was specifically decreased. Based on these findings, we conclude that SHARP-2 is a potential repressor of PEPCK gene expression.