Effects of conditional overexpression of spermidine/spermine N-1-acetyltransferase on polyamine pool dynamics, cell growth, and sensitivity to polyamine analogs

Acetylation of polyamines by spermidine/spermine N-1-acetyltransferase (SSA T) has been implicated in their degradation and/or export out of the cell, The relationship of SSAT to polyamine pool dynamics and cell growth is not yet clearly understood. MCF-7 human breast carcinoma cells were transfected with tetracycline-regulated (Tet-off) SSAT human cDNA or murine gene. Doxy cycline removal for >2 days caused a similar to 20-fold increase in SSAT RN A and a similar to 10-fold increase in enzyme activity. After 4 days, intra cellular putrescine and spermidine pools were markedly lowered, and cell gr owth was inhibited, Growth inhibition could not be prevented with exogenous polyamines due to a previously unrecognized ability of SSAT to rapidly ace tylate influxing polyamines and thereby prevent restoration of the endogeno us pools. Instead, cells accumulated high levels of N-1-acetylspermidine, N -1-acetylspermine, and N-1,N-12-diacetylspermine, a metabolite not previous ly reported in mammalian cells. Doxycycline deprivation before treatment wi th N-1,N-11-diethylnorspermine markedly increased analog induction of SSAT mRNA and activity and enhanced growth sensitivity to the analog by similar to 100-fold. Overall, the findings demonstrate that conditional overexpress ion of SSAT lowers polyamine pools, inhibits cell growth and markedly enhan ces growth sensitivity to certain analogs, The enzyme also plays a remarkab ly efficient role in maintaining polyamine pool homeostasis during challeng es with exogenous polyamines.