Proliferation and morphological transformation of BALB/3T3 cells by a prolonged treatment with sodium orthovanadate

BALB/3T3 mouse embryo cells were used to study the effect of sodium orthovanadate on cell proliferation and morphological transformation. In the presence of the chemical (0.25-1.0 micrograms/ml), the cells continued to proliferate after the cultures were confluent. However, contact-inhibited growth was resumed after removal of the chemical from the culture medium. Continued exposure of the cells to the chemical for 4 wk led to the production of numerous foci consisting of morphologically transformed cells. In contrast, as in vitro transformation assay with a 48-hr treatment protocol followed by 4 wk of incubation without the chemical produced negative results. To test the stability of the transformed foci that were produced on prolonged exposure, we isolated 20 foci with distinctly transformed characteristics from treated cultures and grew them in medium without orthovanadate. 15 isolates gradually reverted to contact-inhibited growth and five maintained the transformed phenotype through ten serial subcultures. The results show that the majority of the transformed foci from the orthovanadate-treated culture failed to maintain transformed characteristics in the absence of the chemical. However, a small fraction of the foci appeared to be altered permanently and exhibited a transformed phenotype in the absence of the chemical.