Figure 3.

The effect of inhibition of endogenous de novo fatty acid synthesis on the uptake of exogenously supplied FAs in OC cells. A2780, OVCAR3 and SKOV3 cells were exposed for 48 hours to increasing concentrations of the FASN inhibitors G28UCM (A) or Fasnall (B) before FA uptake was determined by the QBT™ Fatty Acid Uptake Kit as described in Materials and methods. Obtained dose-response curves (upper panels in A, B) were used to determine optimal drug concentrations (G28UCM: 1.25, 2.5, 10 µM for A2780, OVCAR3 and SKOV3; Fasnall: 25, 50, 100 µM for A2780, OVCAR3 and SKOV3) for subsequent time courses (lower panels in A, B). Comparison of the data in the presence of 5%, 1% or 0% serum reveals that withdrawal of concurrent lipid contained in the serum could not reverse the drug-mediated reduction of uptake of the fluorescent dodecanoic FA analog BODIPY 500/512 C1, C12. Fluorescence signals were normalized to cell numbers that have been determined in parallel by formazan dye cell proliferation assays and were expressed in % of vehicle(0.1% DMSO)-treated control cells grown in the presence of 5% serum. Means ± SD, n = 3. ANOVA followed by Scheffe test, p < 0.05 (*), p < 0.01 (**) and p < 0.001 (***) relative to vehicle-treated cells grown in the presence of the corresponding concentration (5%, 1% or 0%) of serum.