Figure 8. Effect of exosomes on HCC cell behavior.

(A) Transformed cell growth was assessed by determining anchorage-independent growth in soft agar. Cells in culture medium containing 0.4% agarose with or without 15 μg of Hep3B-derived exosomes were plated in a 96-well plate (1000 cells per well) over a base agar layer consisted of culture medium containing 0.6% agarose. After incubation for 7 days, the number of colonies was evaluated fluorometrically and was expressed as fluorescence relative to that in controls without exosomes. Bars represent the mean ± SEM of six separate determinations. *, p < 0.05. (B) Induction of caspase-3/7 activation. Hep3B cells were plated in a 96-well plate (10,000 cells per well) and treated with various concentrations of Hep3B derived exosomes for 24 hours. Caspase-3/7 activity was assessed using a commercial luminometric assay. The data was expressed as relative value of luminescence to control without exosomes. Bars represent the mean ± SEM of three independent experiments. *, p < 0.05. (C) Cell viability assay. Cells (5,000 cells per well) were plated in a 96-well plate with VD medium and incubated with varying concentrations of Hep3B-derived exosomes for 72 hours. Cell viability was assessed using an MTS assay and was expressed as a percentage of control without exosomes. Bars represent the mean ± SEM of 6 separate determinations. *, p < 0.05.