The suppression of cellular proliferation in SV40-transformed 3T3 cells by glucocorticoids

Abstract

Glucocorticosteroids, when added two hours after cell plating to SV40‐transformed, 3T3 mouse fibroblasts in low serum (0.3%v/v), biotin‐supplemented medium, suppress cellular proliferation by 24 hours. While some cell death probably occurs, the growth inhibition is not primarily due to cytotoxicity and cytolysis. This conclusion is supported by the following: (1) both dead and viable cell numbers are suppressed, (2) little cell debris is evident in the medium, and (3) very high concentrations of glucocorticoids do not cause an increase in the dead cell count. Furthermore, this growth suppression, which is specific for glucocorticoids since several non‐glucocorticoid steroids have no inhibitory effect, is not permanent nor irreversible. Removal of the glucocorticoid and replacement with 10% serum restore rapid proliferation. Although higher concentrations (1% and 10%) of serum afford some protection against glucocorticoid inhibition, this protection is not simply a consequence of faster growth rates. SV3T3 cells can be grown in serum‐free medium supplemented with biotin, transferrin, insulin, and epidermal growth factor (EGF). Under these conditions growth rates are comparable to high serum media, yet glucocorticoids are still powerful inhibitors. However, the omission of insulin from serum‐free, glucocorticoid cultures does result in observable cell death and lysis. Flow microfluorometry and autoradiographic studies have determined that glucocorticoid‐inhibited cells are partially blocked in G₁. The proportions of S phase and G₂ + M cells are greatly reduced with an accompanying accumulation of G₁ cells. These results suggest that glucocorticoids regulate a biochemical step(s) in G₁ which is critical for DNA initiation.