We have investigated the possibility that some serum factors might negatively regulate the expression of the insulin-like growth factor-II (IGF-II) gene in 18-54, SF cells. Northern blot analyses indicated that there were three major transcripts (3.8 kb, 1.8 kb, and 1.2 kb) of the IGF-II gene in these cells. We found that incubation of 18-54, SF cells in medium containing very high concentrations (50-100%) of chicken serum greatly inhibited the steady-state level of all three IGF-II mRNA species. In addition, we also found that incubation of 18-54, SF cells in medium containing lower concentrations (10-50%) of chicken serum induced a 3.5 kb IGF-II mRNA. The inhibitory effect of high concentrations of chicken serum on IGF-II mRNA expression was not due to a cytotoxic effect of the serum, because these cells were maintained in 100% chicken serum for up to two weeks without loss of cell viability. The inhibitory effect of chicken serum on IGF-II mRNA was reversible after withdrawal of the serum. Nuclear run-on assays suggested that this negative regulation of IGF-II mRNA in 18-54, SF cells by chicken serum was not the result of transcriptional inhibition. Treatment of 18-54, SF cells that had been previously incubated in 100% chicken serum for 24 h with actinomycin D resulted in a partial restoration of the expression of the 3.8 kb and 1.2 kb IGF-II mRNA in these cells.(ABSTRACT TRUNCATED AT 250 WORDS)