Changes in hepatic differentiation following treatment of rat fetuses with 5-azacytidine.

Rat fetuses of 20 days gestational age were treated in utero with 5-azacytidine. Within 14 to 18 h after treatment several significant changes in the fetal livers were observed, including a dramatic maturation of hepatocyte morphology with little alteration in hematopoietic elements. Assessment of mRNA levels by hybridization to cloned cDNAs, together with other measures of gene expression, established that the change in hepatocyte morphology was associated with strong activation of expression of genes normally activated later in development, including those coding for the liver enzymes tyrosine aminotransferase and phosphoenolcarboxykinase and a gene of unknown specificity that is regulated in liver much like the aminotransferase. Rates of transcription of two of these genes, measured in isolated nuclei, were significantly increased after 5-azacytidine treatment. Expression of alpha-fetoprotein, normally declining during the perinatal period of development, was reactivated following treatment with the drug, while albumin expression was somewhat enhanced. For the most part the changes observed reflect temporal advancement of events normally programmed to occur later in differentiation of the liver. These changes appear to be the consequence of multiple effects of 5-azacytidine, including enhanced gene transcription and stabilization of gene products.