Figure 7.

AC inhibits anchorage-independent growth of breast cancer cells. MDA-MB-453 cells were assayed for their ability to proliferate and form colonies in soft agar. Cells were seeded onto 6 cm dishes in culture medium containing 0.35% low-melting agarose over a 0.7% agarose layer in the presence of AC (40–240 μg/mL) or vehicle control (PBS) and incubated for 3 weeks at 37°C. The numbers of colonies >200 μm in size were counted (at a 40x magnification). Colonies were subsequently stained with p-iodonitrotetrazolium violet (1 mg/mL), and colonies larger than 200 μm were counted. The percentage colony formation was calculated by defining the number of colonies in the absence of AC as 100%. The results are presented as the mean ± SD of three independent assays. *Significant difference in comparison to the control group (P < 0.05).