Regulation of cytochrome P-450c in differentiated and dedifferentiated rat hepatoma cells: role of the Ah receptor.

The induction of cytochrome P 450c mRNA and associated aryl hydrocarbon hydroxylase (AHH) activity is mediated by the Ah receptor in rodent liver and hepatic cells in vitro. In the present study we have investigated the underlying mechanisms responsible for the regulation of AHH activity in differentiated and dedifferentiated variants of the rat hepatoma cell line H4IIEC3. All of the dedifferentiated variants expressed inducible cytochrome P-450c mRNA and AHH activity following treatment with polycyclic aromatic hydrocarbons or the compound 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD). Most of the differentiated derivatives, however, were not inducible for either of these functions. Somatic cell hybridization studies revealed that the differentiated cells were AHH negative due to a defect that corresponded to the Ah receptor D gene product. 5-Azacytidine and sodium butyrate, but not mutagens, reactivated a functional Ah receptor in the differentiated line Fao, indicating that a requisite gene had been silenced by an epigenetic mechanism in this strain. Since many of the 5-azacytidine-induced revertant clones resembled dedifferentiated derivatives with respect to morphology and/or diminished expression of hepatic traits, our results support a correlation between coexpression of the dedifferentiated phenotype and AHH inducibility in these hepatoma cells.