The polyamines putrescine, spermidine, and spermine have been implicated in the regulation of cellular proliferation and differentiation. We have previously demonstrated that spermidine is required for proliferation of murine Friend erythroleukemia (MEL) cells. We have also shown that spermidine is required at a transcriptional level for induction of MEL globin synthesis. Since studies monitoring c-myc expression have suggested that this gene also plays a role in both growth and differentiation, we have monitored the effects of inhibiting ornithine decarboxylase (ODCase) activity and polyamine synthesis on levels of c-myc transcripts. The results demonstrate that the level of c-myc RNA is independent of ODCase inhibition and depletion of intracellular spermidine. More importantly, arrest of MEL proliferation is not associated with detectable changes in c-myc expression, while under these conditions there is a decline in ODCase transcripts. During induction of MEL differentiation with dimethyl sulfoxide (DMSO) and hexamethylene bisacetamide (HMBA), c-myc and ODCase undergo similar changes in patterns of expression. However, although spermidine is required for appearance of the differentiated MEL phenotype, depletion of this polyamine by ODCase inhibition had no detectable effect on the biphasic changes in c-myc RNA observed during MEL differentiation. Thus, these biphasic changes in c-myc expression are not sufficient for induction of the mature phenotype. Finally, these results would indicate that the regulation of c-myc expression during both proliferation and differentiation is independent of ODCase activity and inhibition of proliferation by spermidine depletion.