Dynamic heterogeneity: isolation of murine tumor cell populations enriched for metastatic variants and quantification of the unstable expression of the phenotype

Abstract

Recent studies have indicated that KHT fibrosarcoma or B16 melanoma cell variants capable of forming experimental metastases in the lungs of mice after i.v. injection are created stochastically at high rates (∼10⁻⁵/cell/generation). Expression of this phenotype is unstable and hence expanding populations of tumor cells establish a dynamic equilibrium between a small subpopulation of metastatic variants and a large compartment of nonmetastatic cells. In the present experiments, cell suspensions were prepared from the lungs of mice bearing ‘experimental’ metastases and the tumor cells contained in them were tested for their metastatic efficiency (ME) using the lung colony assay. The ME of the recovered tumor cell populations was found to be a function of the time of metastatic growth in the animal. Tumor cells isolated soon after the initial i.v. injection, i.e. derived from micrometastases, are highly metastatic while populations recovered from macroscopic nodules are similar to parental lines in their ability to colonize the lung. These results are consistent with the prediction of the above ‘dynamic heterogeneity’ model that nascent lung metastases should be composed largely of tumor cells expressing the variant metastatic phenotype, but that the proportion of such variants should decline during growth to the equilibrium (parental population) level. Mathematical analysis of the results indicates that the effective rate of reversion of the variant phenotype is ∼10⁻¹/cell/generation.