The role of selection in progressive neoplastic transformation.

Mathematical modeling indicates that selective growth of cells with biallelic mutations in tumor suppressor genes is the driving force in the development of most human tumors, and that increased mutation rate is not required. Spontaneous neoplastic transformation of cells in culture offers the opportunity for quantitative analysis of all stages of neoplastic progression, the cellular variation that underlies it, and the selective conditions that promote it. Most of the early work on spontaneous transformation was done in primary cultures of mouse embryo cells, but established mouse cell lines have been used more in recent years. The main criteria for transformation have been tumorigenesis in mice, increase in saturation density, and production of discrete, multilayered foci in confluent cell cultures. Spontaneous transformation in NIH 3T3 mouse fibroblasts is efficiently evoked by progressive selection under prolonged contact inhibition at high population density or during multiplication at low population density in suboptimal concentrations or types of serum. In general, it is a multistep process with some stages of progression occurring before there is any visible sign of transformed foci. There is a high degree of heritable heterogeneity in the original NIH 3T3 cell population for susceptibility to transformation. Isolation and expansion of minority susceptible clones from a relatively refractory population exhibit transformation long before the polyclonal parental population does because of the increased proportion of susceptible cells in these clones. There are indications that the selective conditions induce selectable variants. Tumor development in animals and man shares important characteristics with spontaneous transformation in culture, including a major role for selection, but the selective conditions for clonal expansion probably vary with the dynamics of differentiation in each tissue. These considerations support a role for an altered microenvironment (as in the aging process) in selective growth of rogue clones.