Article Figures & Data
Figures
- Fig. 1.
AT-1 tumor sections from untreated (A, E, and I), 3-d castrated (B, F, and J), 3-d treated with ZD6474 (C, G, and K), and 3-d castrated + ZD6474-treated rats (D, H, and L). Bars, 40 μm (A-D and E-H) and 80 μm (I-L). A to D, apoptotic index (active caspase-3). Animals 3-d castrated and 3-d treated with ZD6474 had higher tumor cell apoptosis (arrows, apoptotic tumor cells) compared with those untreated. Animals 3-d castrated + ZD6474-treated had higher tumor cell apoptosis compared with those untreated, 3-d castrated, and 3-d treated with ZD6474. E to H, vascular density (factor VIII–related antigen). Animals 3-d treated with ZD6474 and 3-d castrated + ZD6474-treated had lower vascular density compared with those that are untreated and 3-d castrated. Three-day castrated + ZD6474-treated rats also had lower vascular density compared with those 3-d treated with ZD6474. I to L, pimonidazole staining (tumor hypoxia). Three-day castrated rats had increased fraction of hypoxic cells (brown stained) compared with untreated animals. Three-day castrated + ZD6474-treated rats had increased fraction of hypoxic cells compared with those that are intact, 3-d castrated, and 3-d treated with ZD6474. Three days of treatment with ZD6474 tended to increase the fraction of hypoxic cells compared with untreated animals.
- Fig. 2.
Western blot results of EGFR and VEGFR2 by 7.5% SDS-PAGE under reducing conditions. A, EGFR (Mr 170,000) expression in AT-1 tumor cells. The EGFR expressed in the AT-1 tumor cells gave a somewhat larger band, which has previously been seen in other studies (37). This could possibly be due to differences in glycosylation. B, VEGFR2 (Mr 170,000) expression in AT-1 tumor cells. Rat brain, Mr 170,000, served as a positive control (A and B). The experiment was repeated twice using different rat brain samples and AT-1 tumor cells with similar results (data not shown).
- Fig. 3.
Dose-dependent inhibition of ZD6474 on growth of AT-1 tumor cells measured by fluorometric microculture cytotoxicity assay with an IC50 of 3.1 μmol/L. Abs, absorbance.
Tables
- Table 1.
Effects of castration and ZD6474 treatment in orthotopic AT-1 tumors
Intact day 7* (n = 6) Intact day 10* (n = 15) Castrated day 10* (n = 18) ZD6474 day 10* (n = 13) ZD6474 + castrated day 10* (n = 13) Tumor weight (mg) 15 ± 4.0 64 ± 54† 31 ± 25‡ 22 ± 27‡ 21 ± 20‡ Tumor cell apoptosis (%) 0.44 ± 0.14 0.49 ± 0.28 1.3 ± 0.76‡ 1.3 ± 0.51‡ 1.9 ± 0.59‡§∥ Tumor cell proliferation (%) 38 ± 7.6 32 ± 8.4 26 ± 7.6 35 ± 6.5§ 32 ± 3.8 Volume density of blood vessels (%) 2.5 ± 0.29 3.6 ± 0.80† 3.7 ± 0.66 2.4 ± 0.4‡§ 1.9 ± 0.41‡§∥ Tumor necrosis fraction (%) 0.82 ± 0.93 2.3 ± 2.8 4.8 ± 4.2 3.7 ± 2.9 9.5 ± 4.1‡§∥ Tumor hypoxia fraction (%) NA 56 ± 17 77 ± 16‡ 73 ± 26 91 ± 3.4‡§∥ Abbreviation: NA, not analyzed.
↵* Days after AT-1 tumor cell inoculation. Therapy with castration and/or ZD6474 was given from day 7. For details, see Materials and Methods. Values are presented as means ± SD.
↵† Significantly different than in controls at day 7, P < 0.05.
↵‡ Significantly different than in controls at day 10, P < 0.05.
↵§ Significantly different than in castrated animals at day 10, P < 0.05.
↵∥ Significantly different than in ZD6474-treated animals at day 10, P < 0.05.
- Table 2.
Effects of castration and ZD6474 treatment in the ipsilateral ventral prostate lobe inoculated with AT-1 tumor cells
Intact day 7* (n = 6) Intact day 10* (n = 10-15) Castrated day 10* (n = 14-18) ZD6474 day 10 (n = 8-13) ZD6474 + castrated day 10* (n = 8-13) Ventral prostate weight (both lobes, mg) 251 ± 79 263 ± 59 191 ± 58† 242 ± 58‡ 169 ± 21†§ Apoptosis index (tumor-bearing lobe, %) 0.051 ± 0.0027 0.17 ± 0.05∥ 2.6 ± 0.93† 0.15 ± 0.076‡ 2.4 ± 0.54†§ Proliferation index (tumor-bearing lobe, %) 1.4 ± 0.59 1.9 ± 0.94 2.7 ± 0.71† 1.7 ± 0.78‡ 2.9 ± 0.63†§ Volume density of blood vessels (tumor-bearing lobe, %) 1.4 ± 0.54 1.8 ± 0.32∥ 1.4 ± 0.24† 1.5 ± 0.27† 1.1 ± 0.21†‡§ Apoptosis index (tumor-embedded glands, %) NA 0.76 ± 0.48 2.4 ± 0.98† 1.5 ± 0.72†‡ 2.1 ± 0.50†§ ↵* Days after AT-1 tumor cell inoculation. Therapy with castration and/or ZD6474 was given from day 7. For details, see Materials and Methods. Values are presented as means ± SD.
↵† Significantly different than in controls at day 10, P < 0.05.
↵‡ Significantly different than in castrated animals at day 10, P < 0.05.
↵§ Significantly different than in ZD6474-treated animals at day 10, P < 0.05.
↵∥ Significantly different than in controls at day 7, P < 0.05.
Volume 12, Issue 24
