Effect of N-(n-butyl)-1,3-diaminopropane on polyamine metabolism, cell growth and sensitivity to chloroethylating agents

Abstract

The effects of N-(n-butyl)-1,3-diaminopropane (BDAP) on cell growth and polyamine content were examined in L1210, SV-3T3 and HT-29 cells. In all cases, BDAP was a specific and highly effective inhibitor of spermine synthesis, and spermine levels were greatly suppressed in the presence of 50 [mu]M BDAP. At the same time, there was a parallel increase in spermidine, which equalled or exceeded the fall in spermine so that total polyamine levels were not reduced. Cell growth was not affected in short-term experiments but culture of L1210 cells for 72-144 hr in the presence of BDAP did lead to an effect on growth that was reversed by the addition of spermine. These results suggest that, in the short term, a normal growth rate is maintained by spermidine but that a function or cellular component critically dependent on spermine becomes depleted at longer times. BDAP was a weak inducer of spermidine/spermine-N'-acethyltransferase and this enzyme may be responsible for exretion or degradation of the inhibitor. The reduction of spermine produced by BDAP led to a substantial increase in the activity of S-adenosylmethionine decarboxylase (AdoMetDC) showing that the repression of this enzyme by spermine is greater than the repression by spermidine. Although higher concentrations were required, BDAP was as effective an inhibitor of spermine synthesis as the mechanism-based inhibitor, S-adenosyl-1,12-diamino-3-thio-9-azadodecane (AdoDATAD), and produced similar decreases in spermine and increases in AdoMetDC. Prior treatment of HT-29 human colon carcinoma cells with BDAP increased the killing by chloroethylating agents but to a much smaller extent than the increase brought about by the DNA repair inhibitor, O6-benzylguanine. The effect of BDAP is likely to be due to an increased interaction of chloroethylating drugs with nuclear DNA in the absebce of spermine since BDAP treatment sensitized cells even in the presence of O6-benzylguanine, which prevents repair of these lesions.