The short-term effects of insulin and phorbol esters on the regulation of the albumin gene in rat H4IIE (H4) hepatoma cells were investigated and compared to the expression of a gene known to be inhibited by these agents, phosphoenolpyruvate carboxykinase (PEPCK). Both insulin and phorbol esters inhibited transcription of the albumin gene in a rapid, dose-dependent manner. Within 15 min, albumin transcription was reduced by approx. 80%. The inhibitory effects of insulin were evident at concentrations of insulin as low as 5·10-11 M, suggesting that these effects were mediated through insulin-specific pathways. The ability of both phorbol esters and insulin to inhibit albumin transcription suggests that the negative control of this gene is a stable feature in H4 cells. The effect of phorbol esters to mimic insulin action on the albumin gene, and on several other genes in this cell line, implies that a common pathway may be shared by both insulin and phorbol esters. © 1992.