Expression of the malignant phenotype in rat fibroblasts transfected with the polyomavirus transforming genes.

As a step toward understanding the molecular mechanism of cooperation between viral and cellular genes in oncogenic transformation, we examined various properties of rat cells transfected with the polyomavirus transforming genes without selecting for a neoplastic phenotype. The cell lines displayed a phenotype ranging from nontumorigenic (flat) to fully transformed (tumorigenic). In the established FR3T3 cell line, acquisition of the fully transformed phenotype correlated with effective expression of the polyomavirus middle T (pmt) antigen. Flat cells carrying silent copies of pmt mutated spontaneously to the fully transformed state with a frequency of 2 to 6 X 10(-5) per cell per generation. In unestablished rat fibroblasts, simultaneous transfer of either pmt and small T or pmt and large T in the presence of the neo marker conferred only a partially transformed phenotype to most of the cell lines. The same results were obtained when wild-type genomic DNA was cotransfected with pSV2-neo. The flat transformants progressively acquired properties characteristic of fully transformed cells with passage in culture. However, in contrast to FR3T3 cells, the generation of fully transformed variants from the flat, unestablished fibroblasts was not caused by activation of pmt expression. This indicates that the functions conferred by the large and small T antigens, alone or in combination with each other, cannot substitute for all the functions expressed by the FR3T3 cell line as a result of in vitro establishment. Thus, polyomavirus-mediated transformation may require additional cellular alterations beyond the acquisition of the three viral oncogenes.