Regulation of polyamine synthesis and transport by retinoic acid and epidermal growth factor in cultured adult rat type II pneumocytes

Abstract

During injury of lung epithelial cells, the type II pneumocyte proliferates and differentiates into a type I pneumocyte to restore the epithelium. Polyamines, which constitute a family of small organic polycations, are required for this process of cell repair. Because retinoic acid (RA) and epidermal growth factor (EGF) also are involved, the purpose of this research was to determine their effect on polyamine transport and synthesis in cultured type II pneumocytes. Rat type II pneumocytes were isolated, cultured overnight, and treated with RA and/or EGF for 24 hours. Polyamine transport was determined by [ 3H]spermidine uptake, and polyamine synthesis was assessed by the activity of the initial rate-limiting enzyme ornithine decarboxylase. EGF (100 ng/mL) significantly increased spermidine transport, but RA did not. At low concentrations of spermidine (2 μM), the combined effect of RA and EGF on spermidine transport was additive. Both EGF (25 ng/mL) and RA (1 μM) increased polyamine synthesis, and cotreatment resulted in an additive effect (a fourfold increase over the control). We also found that ornithine decarboxylase activity is greatly diminished in the presence of tyrphostin B56, which is a specific inhibitor for the tyrosine kinase of the EGF receptor, suggesting that polyamine synthesis within the type II pneumocyte may depend on activation of tyrosine kinase of the EGF receptor. These results indicate that RA and EGF increase the availability of polyamines, which may be important in the lung cell repair process.