Colony transformation in C3H 10T1/2 cells: stepwise development of neoplastic change in vitro

This report demonstrates that low passage C3H 10T1/2 cells treated with the carcinogens benzo(a)pyrene or diepoxybutane are transformed morphologically as colonies in as little as 14 d after carcinogen treatment. A transforming dose-response curve is achieved but the frequency of transformation is less than half the expected for 38 d foci, compared on the basis of percent transformants per cell plated. Anchorage-independent cell growth, plating efficiency, doubling time, cell density, and modal chromosomal number were examined from transformed colonies and foci. The data from colony transformants show progressive alteration of these in vitro expressions of neoplastic character with continued subcultivation, consistent with the multistep hypothesis of carcinogenesis. Early in vivo data obtained from one colony-derived transformed cell line show tumorigenesis in irradiated mice within 13 wk of implantation. With continued in vivo passage, tumors were observed in 4 to 6 wk.