The growth of BHK-21/C13 cells in monolayer cultures was inhibited by 6-thioguanosine. Accumulation of putrescine, spermidine, and spermine was inhibited by 6-thioguanosine, and cells incubated in the presence of the drug had a decreased content of polyamines relative to control cells. These effects were more marked for spermidine than for spermine or putrescine. Consequently, the intracellular spermidine:spermine molar ratio was decreased in cells exposed to the drug. Cells, the polyamines of which had been labeled with [3H]putrescine, were incubated in the presence or absence of 6-thioguanosine. More polyamines were lost from cells exposed to the drug than from control cells. The radioactive material excreted was predominantly spermidine, both as its free form and in a conjugated form, even when the cells contained large amounts of labeled spermine. This release of spermidine from BHK-21/C13 cells into the culture medium was a specific response of the cells to alterations in growth potential rather than a consequence of leakage due to cell lysis.