Figure 1. Roc-A inhibits PC-3 cell migration independent of its cytotoxic and anti-proliferative effects.

A. Effect of Roc-A on protein translation. PC3 cells were treated with different doses of Roc-A as indicated. The activities of protein synthesis were monitored by incorporation of ³⁵S-methionine. B. Roc-A decreases cell polarity in PC-3 cells. PC-3 cells were exposed to a gradient of FCS (0-10%) in the presence of 15 nM Roc-A or solvent (DMSO) for 20 h. Examples of polarized (arrow) and unpolarized (arrowhead) cells are indicated. Scale bare = 50 μm. Representative images are shown. C. Quantification of B. At least 230 cells per treatment were analyzed. Results are an average of three independent experiments. Error bars (S.D.) are shown. D. Wound assay. A gap was created in confluent PC-3 cell monolayers and then cells were treated with different concentrations of Roc-A in the absence or presence of zVAD (25 μM for 20 min prior to treatment) for 16 h or treated with 5 μg/ml Mitomycin C (MC) for 1h as described in Material and Methods and then further cultured for 16 h. The gaps before (0 h) and after (16 h) treatment are shown. E. Quantification of the gap closure. The effects of different drugs on cell migration were quantified as percentage of gap closure. Results are an average of three independent experiments. Error bars (S.D.) are shown. F. Analysis of the effects of different drugs on proliferation. PC-3 cells were stained with CFSE as described in Materials and Methods and then seeded and treated as in C. The percentage of proliferative cells was determined and normalized to DMSO treatment. Results are an average of three independent experiments. Error bars (S.D.) are shown. G. Analysis of the effects of different drugs on cell viability. The percentage of viable cells was determined as AnxV⁻/7aad⁻ cells 20 h after drug treatment. Results are an average of three independent experiments. Error bars (S.D.) are shown.