A RAA > 1 indicates potential clinical activity. Results Single agent antiproliferative activity of FWGE in human cancer cell lines The antiproliferative activity of a 96 hour continuous exposure GDC-0994 ic50 to FWGE was evaluated in a large panel of human tumor cell lines using the SRB-assay. IC50-values were calculated

using the Hill equation and the obtained data from at least three independent experiments were summarized as a mean graph (Figure 1). IC50 of FWGE ranged from 0.038 mg/ml to 0.7 mg/ml with a median IC50 of 0.33 mg/ml. Figure 1 Illustration of IC 50 of FWGE as a mean graph. IC50 of at least 3 independent experiments per cell line were averaged and summarized as a mean graph for better comparison of the different activity. The average IC50 is 0.33 mg/ml. The highest activity of FWGE was found on neuroblastoma and ovarian cancer cell lines. It’s interesting to note that the IC50-values of the 8 human CRC cell lines included in this screen range close to the average IC50. Notably, the estimated peak plasma concentration after the

oral intake of a standard dose of 9 g/day FWGE in patients is 0.5-1 mg/ml [7]. Considering this peak plasma concentration and the observed IC50 in our cell line screen, the calculated RAA is at least 1 or higher which could indicate potential clinical activity. The highest BX-795 cost activity of FWGE was found in neuroblastoma cell lines with an average IC50 of 0.042 mg/ml (RAA ≈ 12-24). Of note, the 8 colon cancer cell lines included in this screen had a very narrow IC50 range varying from 0.3 mg/ml to 0.54 mg/ml yielding in a RAA of 1.7-3.3 (Figure 1). Detection of the mode of cell death induced by FWGE in a panel of cell lines In order to distinguish the mode of cell death induced by FWGE we treated a representative panel of human cancer cell lines with an IC90 of FWGE for 48 h. Subsequent to treatment, floating cells were harvested and an DNA gel electrophoresis was performed. Clearly, in all treated

cell lines the typical 180 bp DNA laddering structure indicative for specific DNA degradation during the process of apoptosis could be detected (Figure 2). Figure 2 Induction of apoptosis by FWGE. learn more A representative panel of human tumor cell lines was treated with an IC90 of FWGE for 48 h and floating cells were harvested by centrifugation for DNA extraction. DNA was seperated by DNA gel electrophoresis and stained with ethidium bromide subsequently. Typical DNA laddering indicative for apoptosis was visualized by UV light illumination. Combination of FWGE with 5-FU, Oxaliplatin and Irinotecan in human colon cancer cell lines The combined drug effect of a parallel exposure to FWGE and either 5-FU, irinotecan or oxaliplatin was assessed in a panel of 8 colon cancer cell lines. The mode of drug interaction was analyzed by the method of Drewinko and the data summarized in table 1.