An epithelioid clone of Chinese hamster (CHO) cells which is spontaneously transformed was exposed to the mutagen N-methyl-N-Nitro-N-Nitrosoguanidine (MNNG), and a fibroblastic variant, clone CHO-F2, was isolated. This clone is partially reverted in several of the in vitro properties characteristic of transformed cells. When compared to wild type CHO, CHO-F2 has a longer doubling time, a lower saturation density and less piling up at high cell density, and a higher serum requirement. CHO-F2 also elaborates less plasminogen activator and has more abundant microtubules and actin cables. On the other hand, both CHO and CHO-F2 grow in agar suspension (although CHO-F2 grows with a lower efficiency), both lack detectable LETS protein, and both are tumorigenic in nude mice. Thus, expression of the individual properties frequently associated with transformation and tumorigenicity can be dissociated. The most critical biochemical change in CHO-F2 appears to be an increase in the intracellular level of cyclic AMP, when compared to CHO, and several growth and morphological properties of CHO-F2 resemble those induced in wild type CHO exposed to exogenous dibutyryl cyclic AMP. The role of cyclic AMP in expression of the transformed phenotype and the significance of individual in vitro parameters of transformation with respect to tumorigenicity are discussed.
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