Role of polyamines in differentiation of 3T3-L1 fibroblasts into adipocytes.

The role of polyamines in the differentiation of 3T3-L1 fibroblasts into adipose cells was studied. This conversion was blocked by the addition of alpha-difluoromethylornithine, an enzyme-activated irreversible inhibitor of ornithine decarboxylase, which prevented a rise in spermidine content in the differentiating cells. The inhibition of differentiation could be overcome completely by the provision of exogenous putrescine, spermidine, or spermine. Partial reversal could be produced by exposure to nonphysiological homologues of the natural polyamines such as 1,3-diaminopropane, 1,5-diaminopentane, and sym-norspermine. Reversal of the inhibition of differentiation by exogenous polyamines required a period of exposure to the amines, indicating that the lack of differentiation is not due simply to an obligatory role for polyamines in the biosynthesis of lipids. These results indicate that spermidine is required for the differentiation, but spermidine alone was not able to replace insulin and 1-methyl-3-isobutylxanthine in stimulating conversion to adipocytes. Therefore spermidine appears to be necessary but not sufficient for differentiation to occur. Finally, the elevation of spermidine content that occurs during the conversion of fibroblasts to adipocytes did not correlate with an increased activity of the polyamine biosynthetic enzymes. This implies that the increase must be regulated by changes in the rate of degradation or excretion of the polyamines.