Developmental activation of the H-2K gene is correlated with an increase in DNA methylation

Embryonal carcinoma cell lines present strong developmental analogies with early embryonic cells. Previous studies have shown that treatment of F9 teratocarcinoma cells with retinoic acid induces the expression of the classical transplantation antigens which are indispensable for effective interactions between cells. In contrast to several genes that were analyzed, all of which were highly and heterogeneously methylated in F9 cells, the H-2K gene was poorly methylated if at all. Activation of the H-2K gene upon differentiation of F9 cells was accompanied by an increase in DNA methylation. While increased methylation of the H-2K gene in one of the two homologous chromosomes was correlated with a low level of expression, increased methylation in both chromosomes was associated with a high level of expression. Treatment of differentiated F9 cells with 5-azacytidine resulted in inhibition of DNA methylation and a concomitant repression of the H-2K gene expression. Thus, in contrast to the many examples of an inverse correlation between the level of gene methylation and its transcriptional activity, expression of the H-2K gene is directly correlated with the extent of methylation. This finding offers an explanation whereby the hypomethylation observed in tumor cells may be responsible for the establishment and maintenance of a malignant state of growth.