Effects of Dexamethasone and Anabolic Agents on Proliferation and Protein Synthesis and Degradation in C2C12 Myogenic Cells

The objective of this experiment was to determine the dose response of dexamethasone (DEX) on C2C12 myogenic cells and to examine the effects of the anabolic compounds estradiol (E), testosterone (T), and dihydrotestosterone (D) alone and in combination with DEX on proliferation and protein turnover in cultured C2C12 myogenic cells. In the first study, cells were treated with seven concentrations (0, 25, 50, 75, 100, 150, or 200 nM) of DEX in medium with or without 5% horse serum (HS) for the determination of protein synthesis and degradation, and six concentrations (0, 50, 100, 150, 200, or 250 nM) of DEX in medium with 5% fetal bovine serum for cell proliferation measurements. Proliferation of myoblasts decreased (P < .05) with DEX. As DEX concentration increased, protein degradation in myotubes increased (P < .05) up to 100 nM, then declined. Protein synthesis decreased linearly (P < .01) as DEX concentration increased. The presence of HS in the medium decreased (P < .01) protein degradation by 32% as compared with no HS and increased (P < .05) protein synthesis. In the second study, cells were treated with E, T, or D at four concentrations (0, 100, 500, or 1,000 nM) in medium containing 0 or 100 nM DEX. Cells were assayed for protein synthesis or protein degradation. Synthesis decreased (P < .01) and degradation increased (P < .01) with DEX. No differences (P > .05) were found between E, T, or D hormone treatments or concentrations. To measure proliferation, myoblasts were treated 1 d after plating with the same anabolic hormone treatments in medium containing 0 to 100 nM DEX. Cells were grown to confluence and assayed for proliferation. Proliferation decreased (P < .01) in the presence of DEX in each treatment compared with controls. Cells treated with E had significantly lower (P < .05) proliferation rates than cells treated with T and D. The presence of concentrations of DEX at 100 nM inhibited proliferation and protein synthesis and increased protein degradation. Anabolic agents at pharmacological doses do not inhibit the DEX effects on C2C12 myogenic cells, nor do they directly affect proliferation or protein turnover.