Development of a System of Study Gene Activation in Mammalian Cells Treated with Bis (2-Chloroethyl) Sulfide

摘要

It is known that treatment of mouse S49 thymic lymphoma cells with N-acetoxy-2-acetylamino-fluorene (NA-AAF) or ethylnitrosourea (ENU) activate two quiescent genes, metallothionein I (MT-I) and metallothionein II (MT-II) (Annual Summary Report, 12/1/86). We reported that treatment with NA-AAF and ENU resulted in a 10-30 fold increase in cadmium-resistant variants and that about 65% of these variants made either MT-I, MT-II or both. Among other findings, we reported that the DNA methylation patterns (As measured by Hpa II digestion and Southern blotting), changed in lines which were MT positive. Using restriction endonuclease digestion and Southern blotting, we have not detected any insertions, deletions, rearrangements, or amplifications associated with the activation of either MT gene. We are now in the process of analyzing metallothionein expression in these Cdr variants. Keywords: Cultured mouse cells; Gene activation; Sulfur mustard; Alkylating agents; Metallothionein; Nucleic acid hybridization; Gene rearrangement; DNA Methylation. (JES)