Gastrin induction of mRNA expression in rat colonic epithelium in vitro

A newly developed system of isolated rat colonic epithelial cells was utilized for a comprehensive study of protein synthesis influenced by gastrin. We found that synthetic human gastrin (0.01-100 nM) increased the incorporation of [35S]methionine into proteins within 2 hours. Peak incorporation was observed with 10 nM gastrin to more than two-fold above maintenance levels. Actinomycin-D (10 micrograms/ml) inhibited the stimulated increases in total protein synthesis indicating that the peptide's trophic effect was mediated by the synthesis of new mRNA species. The effect of gastrin was comparably stronger than the one induced by the mitogen bombesin (1 nM). However, bombesin, a neuromodulator of gastrin release, did not produce an additive effect beyond that of gastrin on total protein synthesis. Gastrin stimulated the synthesis of many polypeptides resolved on two-dimensional polyacrylamide gel, an indicator of gastrin's influence on the expression of various mRNA species. Some of these polypeptides may be used as markers in investigating colonic epithelial response to gastrin.