Phenobarbital induction of cytochrome P-450 in normal and preneoplastic rat liver: comparison of enzyme and mRNA expression as detected by immunohistochemistry and in situ hybridization.

The expression levels of the major phenobarbital-inducible cytochromes P-450 were analyzed immunohistochemically in liver sections obtained from rats treated sequentially with diethylnitrosamine and phenobarbital. In parallel sections expression of the corresponding messenger RNAs was investigated by in situ hybridization using an appropriate 35S-labeled cDNA probe. A phenobarbital-mediated increase in enzyme protein and a concomitant enhancement of the corresponding mRNA was seen in hepatocytes of the centrilobular areas of the liver acinus. A comparable increase in P-450 protein and mRNA expression was detected in large carcinogen-induced nodules present in animals maintained on phenobarbital until killing. This result was confirmed by Northern analysis of nodular RNA. No such induction was seen in liver nodules obtained from animals which were withdrawn from the phenobarbital-containing diet 10 days prior to killing. These findings demonstrate that phenobarbital induction of cytochrome P-450 enzymes occurs in neoplastic liver tissue as it does in normal liver and, in addition, suggest that the increase in the levels of the major phenobarbital-inducible P-450s in normal and neoplastic liver is mediated by an enhancement of the steady-state concentrations of their mRNAs.