Conditioned medium from cultured rat hepatocytes completely blocks induction of glutamine synthetase by dexamethasone in several rat liver epithelial cell lines.

A variant RL-ET-1G of a rat liver epithelial cell line (RL-ET-1) characterized by a very high inducibility for glutamine synthetase (GS) in response to dexamethasone was established by cultivation in glutamine-free, glutamate-supplemented culture medium. Using this cell line, conditioned medium produced by periportal hepatocytes in primary culture was found to suppress this induction, acting with a lag-phase of about 8 h irrespective whether the GS activity was basal or preinduced. Analysis of the response of several epithelial cell lines to the conditioned medium showed a reciprocal relationship between the dexamethasone-dependent induction and the residual activity after exposure to the conditioned medium, indicating that a hypothetical factor in the conditioned medium was interfering with the induction process but not with the basal GS level of these cells. Careful analysis revealed that the effect of the conditioned medium was neither due to deficiency of a component used up by the hepatocytes, nor due to glutamine or ammonia, both of which affected GS activity at concentrations above 0.5 mmol/L. The hypothetical factor was found to be quite small (molecular mass range 100-500 Da), heat and acid stable, as well as highly water soluble. Most interestingly, the conditioned medium did not suppress GS induction in astroglial cells and in the two hepatoma cell lines C2 and FAO, but strongly diminished the spontaneous induction of GS in cocultured pig hepatocytes, suggesting that the hypothetical factor acts primarily on normal nontransformed liver-derived cell populations.