The effects of epidermal growth factor, interleukin-1, and phenytoin, alone and in combination, on C19 steroid conversions in fibroblasts.

Abstract

The formation of the biologically active metabolite 5 alpha-dihydrotestosterone (DHT) from testosterone in response to phenytoin (Ph), interleukin-1 (IL-1), and epidermal growth factor (EGF) was investigated. The androgen DHT stimulates matrix synthesis in connective tissue and bone. Duplicate incubations were performed with confluent human gingival fibroblasts, 14C-testosterone, and optimal stimulatory concentrations of IL-1 (5 IU/ml), EGF (10 ng/ml), Ph (5 micrograms/ml), Ph + EGF, and Ph + IL-1 respectively for 24 hours in Eagle's MEM at 37 degrees C. The medium was then analyzed for radioactive metabolites. Similar incubations were performed with human gingival tissue using 14C-4-androstenedione as substrate in the presence or absence of EGF, Ph, and EGF + Ph. In the cell lines studied, EGF stimulated DHT and 4-androstenedione synthesis by 20% (n = 5; P < 0.01; Wilcoxon signed rank statistic for paired observations). IL-1 stimulated DHT and 4-androstenedione synthesis by 2-fold (n = 6; P < 0.01). Ph stimulated DHT and 4-androstenedione synthesis by 2-fold increases (n = 3; P < 0.01). Combinations of phenytoin and EGF stimulated DHT and 4-androstenedione synthesis by 33% and 37% greater than the effect of phenytoin alone (n = 3; P < 0.01). Combinations of Ph and IL-1 caused a 45% increase in the amount of DHT formed and a 66% increase in 4-androstenedione when compared to the effect of phenytoin alone (n = 3; P < 0.01). 14C-4-androstenedione was converted to DHT and testosterone by human gingival tissue. There were 2-fold, 4-fold, and 2.5-fold increases in DHT synthesis and 5-fold, 2-fold, and 6-fold increases in the formation of testosterone in response to EGF, Ph, and EGF + Ph respectively (n = 3; P < 0.01). EGF and IL-1 present in inflammatory exudate may have implications on phenytoin-induced overgrowth via the steroid metabolic pathway.