Figure 1. The effect of DHA, EPA and AA on the viability of hepatocellular carcinoma cells.

Equal numbers of human HCC cells (Hep3B, HepG2 and Huh-7) were seeded onto the 96-well plates (1 × 10⁴ per well) and cultured in the medium supplemented with 10% fetal bovine serum for 24 hr. The cells were washed twice with PBS and maintained in serum-free medium for 24 hr before treatment with PUFAs. The cells were treated with DHA, EPA or AA at increasing concentrations (1–100 µM) for indicated time periods (12–72 hours). (A) The effect of DHA, EPA or AA on the viability of Hep3B cells (30 µM, 24 hours). The cell viability was determined using WST-1 assay. The data are presented as mean ± SD of six independent experiments. (B) Dose-dependent reduction of cell viability by DHA and EPA in Hep3B, Huh7 and HepG2. The cells were treated with 1–100 µM individual PUFAs for 24 hours. The cell growth was determined using WST-1 assay. The data are presented as mean ± SD of six independent experiments. (C) DHA induces the cleavage of caspase-3, caspase-9 and PARP and release of cytochrome c in Hep3B cells. (a) DHA induces the cleavage of caspase-3, caspase-9 and PARP. Hep3B cells were treated with DHA (30 µM) for 24 hr and the cell lysates were obtained for Western blot analysis using antibodies against PARP, caspase-3 and caspase-9. (b) DHA induces the release of cytochrome c in Hep3B cells. The levels of cytochrome c in the cytosolic and mitochondrial fractions were determined by western blotting analysis. EPA was not utilized in these experiments.