Expression of a 32 kDa protein in rat mammary tumors induced by anti-benzo[c]phenanthrene-3,4-diol-1,2-epoxide

Racemic anti-benzo[c]phenanthrene-3,4-diol-1,2-epoxide (BcPDE) is a powerful rat mammary carcinogen and a metabolite of benzo[c]phenanthrene, a polynuclear aromatic hydrocarbon found in the environment. In elucidating potential molecular mechanisms that may play a role in the development of BcPDE-induced rat mammary tumors, we have identified a 32 kDa protein in 16 of 26 tumors analyzed but in only 1 of the 15 normal mammary tissues that were examined. The 32 kDa protein was identified with antibodies to Ets, which also recognized the 55 kDa Ets-1 protein that was expressed at similar levels in normal mammary tissues. The expression of the 32 kDa protein was also observed in mammary tumor-derived cell lines of both rat and human origin and in human melanoma, but not in normal human keratinocytes or rat fibroblast cell lines. Further characterization via 20 gels revealed that the protein exhibits a PI of 5.5. Southwestern analysis using Ets-1 target sequence revealed binding of the 55 kDa Ets-1 but not of the newly identified 32 kDa protein. Overall, the preferential expression of the 32 kDa protein in mammary tumor tissues may serve as a biomarker to follow the development of this tumor type.