A number of human esophageal (3) and bronchial (10) cancers have been characterized clinically and by their histopathology. These tumors have been investigated for the persistence of human adenoviral DNA sequences. The polymerase chain reaction (PCR) and Southern transfer hybridization (SBH) techniques have been applied. All analyses have consistently yielded negative results. These findings are discussed in the light of comparisons to the Ad12 hamster tumor system in which tumor cell or transformed cell revertants can lose the integrated Ad12 DNA sequences, but retain the oncogenic phenotype, when reinjected into hamsters. Ad12-transformed cells and Ad12-induced tumor cells have previously been shown to exhibit altered cellular methylation and transcription patterns. In one of the revertants, which has lost all Ad12 DNA sequences, changes in cellular DNA methylation patterns are also maintained. Since in the hamster tumor system the loss of Ad12 DNA sequences is still compatible with the oncogenic phenotype, the possibility exists that human tumors, though themselves devoid of viral DNA sequences, could have had cells as precursors which originally carried integrated adenoviral DNA sequences.