Article Figures & Data
Figures
- Fig. 1.
The effect of AS 4625 and control 4626 on cancer cell growth (A), protein expression (B), and VEGF secretion (C). A, dose-response effect of 4625 and 4626 on the soft agar growth of GEO colon and ZR-75-1 breast cancer cells. Data represent means and SE of three different experiments, each performed in triplicate. Cells were counted as described in “Materials and Methods.” B, effect of 4625 and 4626 on bcl-2, bcl-xL, and actin protein expression. Western blotting analysis of protein expression in GEO and ZR-75-1 cells grown in monolayer, untreated or treated for 4 days with 1 μm 4625 or 4626. Cell lysates were processed as described in “Materials and Methods.” C, VEGF secretion in the CM collected from ZR-75-1 cells, untreated or treated for 4 days with either 0.5 μm 4625 or 1 μm 4626, as described in “Materials and Methods.” Data represent the average (±SD) of two different experiments each performed in triplicate.
- Fig. 2.
The effect of either 4625 (A) or 4626 (B) in combination with AS-PKAI and ZD1839 on the soft agar growth of GEO and ZR-75-1 cells. A, 0.5 μm 4625 (a–c), 0.25 μm 4625 (d–f), 0.1 μm AS-PKAI (a, c, d, and f), and 0.1 μm ZD1839 (b, c, e, and f). B, 0.5 μm 4626 (a–f), 0.1 μm AS-PKAI (a, c, d, and f), and 0.1 μm ZD1839 (b, c, e, and f). Data are expressed as a percentage of colony formation inhibition compared with untreated control cells. The first bar of each couple shows the individual effects of each drug when used alone (represented as stacked bars). Thus, the total height of these stacked bars also represents the expected total inhibition, if drugs have an additive effect. The second bar of each couple (black bar) shows the effect obtained when the drugs were actually used in combination. Therefore, the comparison between the height of the first bar and that of the second bar of each couple shows whether a supra-additive effect is obtained and the magnitude of such effect. The data represent means and SE of triplicate determination of at least two experiments.
- Fig. 3.
The effect of either 4625 (A) or 4626 (B) in combination with AS-PKAI and ZD1839 on apoptosis of GEO and ZR-75-1 cells. A, 1 μm 4625 (a–c), 0.5 μm 4625 (d–f), 0.5 μm AS-PKAI (a, c, d, and f), and 0.5 μm ZD1839 (b, c, e, and f). B, 1 μm 4626 (a–f), 0.5 μm AS-PKAI (a, c, d, and f), and 0.5 μm ZD1839 (b, c, e, and f). Treatments were carried out as described in “Materials and Methods.” Data are expressed as apoptotic index, which represents the ratio between the absorbance of treated cells and that of untreated cells, normalized for the same number of cells. Therefore, results for each treatment are presented relative to control untreated cells, referred to as 1. The percentage of apoptotic cells is ∼4% in untreated GEO cells and ∼6% in untreated ZR-75-1 cells, as determined by flow cytometry. The data represent means and SE of duplicate determination of at least two experiments.
- Fig. 4.
The effect of the treatment on the growth (A) and protein expression (B) of GEO tumor xenografts in nude mice. A, the schedule of each single agent, alone or in combination, is described in “Materials and Methods.” The doses of each drug, used alone or in combination, were: (a) 20 mg/kg 4625; (b) 20 mg/kg 4626; (c) 10 mg/kg AS-PKAI; and (d) 150 mg/kg ZD1839. B, Western blot analysis was performed on total lysates from tumor specimens of two mice sacrificed at day 25, as described in “Materials and Methods.”
Volume 9, Issue 2
