A mammalian cell culture system using normal, diploid Syrian hamster embryo fibroblasts was used as a model to study the ability of diethylstilbestrol (DES) and related compounds to induce neoplastic transformation. Like benzo(a)pyrene, a known chemical carcinogen, DES (0.01 to 10 micrograms/ml) induces morphological transformation of Syrian hamster embryo fibroblasts in vitro; the transformed colonies were indistinguishable from colonies of benzo(a)pyrene-transformed cells. The morphologically transformed colonies derived from either benzo(a)pyrene- or DES-treated cells were tumorigenic when injected into newborn hamsters. At doses of 0.01 to 1 microgram/ml for 48 hr, DES did not inhibit or stimulate cell proliferation. Structural analogs of DES were also tested in the Syrian hamster embryo transformation system. Like DES, tetrafluorodiethylstilbestrol, dimethylstilbestrol, and cis,cis-dienestrol morphologically transformed Syrian hamster embryo cells. The transformation frequency of dimethylstilbestrol was much less than that of DES, tetrafluorodiethylstilbestrol, or cis,cis-dienestrol. Hexestrol, dimethoxydiethylstilbestrol, and trans,trans-dienestrol did not transform these cells. No correlation could be demonstrated between reported estrogenic potency of the compounds and their cell-transforming capacity. Structure-activity relationships developed for these chemicals suggest that metabolism via specific pathways plays a role in DES-induced cell transformation.