The morphological oncogenic signature. Reorganization of epithelial cytoarchitecture and metabolic regulation by tumor promoters and by transformation.

The dramatic changes in morphology induced by nanomolar doses of tumor-promoting agents, especially in epithelial cells, have been noted previously (Driedger and Blumberg, 1980; Rifkin et al., 1979; Croop et al., 1980; Phaire-Washington et al., 1980; Ohuchi and Levine, 1980; Ojakian, 1981; Fey and Penman, 1984). This chapter shows the effect of the tumor promoter TPA on the underlying skeletal framework, which is involved in the maintenance of both cell and epithelial tissue morphology. It should be emphasized, however, that similar results are obtained for all the tumor promoters as well as for the complete, ultimate carcinogens examined so far. The organization of the cytoskeletal elements involved in these morphological changes is faithfully retained during the fractionation procedure employed here, as is evident from SEM and TEM analysis of Triton-extracted cells. A number of promoting agents have been compared, and the degree of disorganization viewed in these whole mounts appears to parallel the potency of the promoting agents as measured by other assays (Fey and Penman, 1984). Also, the inactive analogues of phorbol ester have no effect on cell structure (Rifkin et al., 1979; Ojakian, 1981; Fey and Penman, 1984). We suggest that the effect of TPA on the cytoskeleton occurs early as compared with many of the commonly studied biochemical responses and may indeed underlie many of the previously described cellular response to promoting agents, such as mitogenic stimulation. TPA-induced alterations in NM-IF scaffold occur in the absence of both protein and RNA synthesis (Fey and Penman, 1984). By contrast, plasminogen activator, stimulated by TPA (Wigler and Weinstein, 1976), is completely blocked by pretreatment with both cycloheximide and actinomycin D (Weinstein et al., 1977; Ojakian, 1981). Ornithine decarboxylase, another enzyme that is rapidly induced by tumor promoters, is inhibited by both cycloheximide and actinomycin D in the presence of TPA (O'Brien, 1976). Thus two of the early biochemical markers for tumor-promoter activity are separable from the induction of cytoskeletal alterations by TPA. One of the most striking features of the response to promoting agents is the adoption of the transformed phenotype, in which cells lose growth control and cease being organized into meaningful tissue structure. The alteration of desmosomal and junctional associations and the concomitant change in cytokeratin organization are clearly related to the breakdown of epithelial organization. The phenotype is completely reversible although it takes about 3 days for the mode line to reestablish normal morphology (data not shown).(ABSTRACT TRUNCATED AT 400 WORDS)