Dissociation of contact-noninhibition in vitro and tumorigenicity in vivo.

For cell lines, the correlation of the in vitro expression of contact-inhibition to the in vivo expression of the nontumorigenic phenotype, and the correlation of the expression of contact-noninhibition (i.e., focus formation) to the expression of the tumorigenic phenotype are commonly used as a means to identify, in vitro, cells which presumably have undergone malignant transformation such that, if tested, they would grow as tumors in vivo. In this report we show that while this correlation is true for contact-noninhibited transformants induced by benzo(a)pyrene (BP), a DNA mutating and demethylating agent, it is not true for contact-noninhibited transformants induced by 5-azacytidine (AZC), a DNA demethylating agent which does not have mutagenic activity. The in vitro treatment of a contact-inhibited cell line with 5-azacytidine (AZC) results in the expression of contact-noninhibited transformants, which, in vitro quantitatively and qualitatively similar to those induced by BP but unlike BP, treatment with AZC does not result in the in vivo expression of tumorigenicity.