Figure 1

P(122-131) promotes anti-tumor phenotypes in DU145 and LNCaP cells and inhibits angiogenesis in vivo. (A) Number of adherent DU145 and LNCaP cells in the presence of increasing concentrations of P(122-131). An equal number of DU145 or LNCaP cells was incubated with increasing concentrations of P(122-131) for 30 min before seeding. After a 10-min incubation period, adherent cells were measured by the crystal violet assay. (B) Soft agar growth assays showing anchorage-independent proliferation. An equal number of DU145 or LNCaP cells was resuspended in growth medium containing 10% FBS, 0.3% agar, and increasing concentrations of P(122-131), and seeded onto the bottom agar, which consisted of growth medium containing 10% FBS and 0.8% agar. The top agar was allowed to solidify, and standard growth media supplemented with peptide was added to each well. The cells were incubated 12 days, after which cell colonies larger than 50 μm were quantified by counting the entire area of each well. (C) Migration of cells through Transwell filters. The lower compartment of Transwell filters (8 μm pores) was filled with growth media containing 2.5% FBS, 0.5% BSA, and increasing concentrations of P(122-131). An equal number of DU145 or LNCaP cells was resuspended in growth medium containing 2.5% FBS and 0.5% BSA, and transferred into Transwell inserts. Cells that successfully migrated through the filter pores, were fixed, stained and quantified by counting the entire area of each filter. (D) Effect of P(122-131) on angiogenesis, as measured by the chicken embryo CAM assay. An 1 cm² area of chicken embryo CAM, restricted by a silicon ring, was incubated with increasing concentrations of P(122-131). 48 h later total vessels length was quantified as described in Materials and Methods. Results are mean values ± SE from at least 3 independent experiments.