Clonal dynamics of progressive neoplastic transformation

In a recent study, we found that newly isolated clones of NIH 3T3 mouse cells undergo neoplastic transformation more readily than uncloned cultures from which they were derived. After eleven low-density passages (LDPs), most of the 29 clones produced lightly stained early-stage transformed foci when grown to confluence in a primary assay for transformation, and one of them consistently produced a few tiny dense foci, In the present work, six of the clones were kept in LDPs for 56 passages and assayed for focus formation at confluence at six passage levels. The clone that produced tiny dense foci switched to light foci during the LDPs, four others produced light foci at different passage levels, and one progressed from light to dense foci after the last passage. By contrast, all the clones progressed to dense focus formation in five or fewer serial repetitions of the assay at confluence. Because all but one of the clones underwent about half as many total divisions at each LDP as they did when grown to the stationary state at confluence, the latter is more efficient in eliciting progression than the exponential growth of the LDPs, Extension of the period at confluence of uncloned cultures results in the appearance of dense foci within light foci, Because the latter are localized clonal populations, the intrafocal progression reinforces the conclusion that clonal expansion favors transformation. We discuss the significance of these results for the clonal origin of human cancer and the increased incidence of cancer with age.