Induction of prolactin-deficient variants of GH3 rat pituitary tumor cells by ethyl methanesulfonate: reversion by 5-azacytidine, a DNA methylation inhibitor

Abstract

GH3 cells are a rat pituitary tumor line expressing two pituitary peptide hormones, prolactin (rPRL) and growth hormone. Recently, it was found that the DNA alkylating agent ethyl methanesulfonate can induce the appearance of rPRL-deficient GH3 cell variants at a high frequency (ca. 20-30%). As shown here, such variants cannot be induced at high frequency by irradiation of wild-type GH3 cells with ultraviolet light, indicating that the effect may be specific to treatment with alkylating agents. Furthermore, the DNA methylation inhibitor 5-azacytidine reverted an ethyl methanesulfonate-induced rPRL-deficient variant into rPRL-expressing cells at high frequency (ca. 50%). The revertants were stable for at least 30-35 generations. These results support the hypothesis that the alkylating agent may promote the specific methylation of the rPRL gene or a gene regulating its activity, either one of which leads to inactivation of expression of the rPRL gene in GH3 cells.