Inhibition by retinoids of neoplastic transformation in vitro: cellular and biochemical mechanisms

We have demonstrated that non-toxic concentrations of retinoids can cause a dose-dependent inhibition of 3-methylcholanthrene-induced transformation of C3H/10T1/2 cells. On removal of the retinoid, transformed foci appear after a latent period of about four weeks at the same frequency as observed in controls treated with carcinogen only. Reasoning that this activity is compatible with the stabilization of the carcinogen-initiated state, we have succeeded in isolating from carcinogen-treated cultures a cell line which in the presence of retinyl acetate is similar to the parental 10T1/2 cells, but without retinyl acetate transforms at a high frequency after a latent period of about four weeks. Retinyl acetate treatment of this cell line (INIT/10T1/2) and the parental 10T1/2 cells induces an ultra-normal phenotype. In retinoid-deprived INIT/10T1/2 cells, the first sign of transformation (an increased thymidine-labelling index) occurs 16 days after retinyl acetate removal. We have detected by two-dimensional electrophoresis that concomitant with this there is an increase in phosphorylation of a protein of Mr 34 000 (34K) which may be associated with the cytoskeleton. This phosphoprotein has been found in all transformed lines examined. A second phosphoprotein, of about 38K, has also been detected in transformed cells. Retinyl acetate treatment of transformed cells alters the isoelectric point of this protein, a change compatible with decreased phosphorylation. Alkali-resistant phosphorylation, presumably on tyrosine, has been found on a second 34K protein of transformed cells. Retinyl acetate treatment specifically decreases this phosphorylation. Mechanisms for the altered tyrosine phosphorylation induced by retinyl acetate are as yet unresolved, but the decrease could be due to altered levels of substrate, kinase or phosphorylase. In view of the apparent role of tyrosine kinases as mediators of growth factors and as oncogene products, we consider the activity of retinoids as modulators of tyrosine phosphorylation to be of great potential significance.