Article Figures & Data
Figures
- Fig. 1.
Effects of SAHA on prostate cancer cells in vitro. A, cells (LNCaP, PC-3, or TSU-Pr1) were treated with SAHA at the following concentrations: 0 (control),▪ ; 2.5 μm SAHA, ♦; 5 μm SAHA, •; and 7.5 μm SAHA, ▴. Cells were counted at the indicated times using a hemocytometer, and the number of viable cells was determined by trypan blue exclusion. The top panels show the number of viable cells present at the time points indicated, whereas the bottom panels show the number of dead cells as a percentage of the total cells present in each culture. Data are presented as the mean ± SE of triplicate values from a typical experiment. B, LNCaP cells (5 × 106) were treated with the indicated concentrations of SAHA for 3 h. Cells were harvested, and histones were prepared as described in “Materials and Methods.” Histone acetylation was detected by Western blot using antibodies against acetylated H3 and H4. The top panel contains acetylated histone H3, the middle panel contains acetylated histone H4, and the bottom panel shows a Coomassie blue-stained polyacrylamide gel of the total histones extracted from the cells.
- Fig. 2.
s.c. growth of the CWR22 prostate cancer xenograft in nude mice treated with daily i.p. injection of vehicle alone or SAHA (25, 50, or 100 mg/kg). Data are presented as the mean tumor volume ± SE of the surviving animals in each treatment group. All groups contained eight or nine mice at each time point, except for the group receiving 100 mg/kg/day SAHA, in which five animals died by the end of the treatment period. Treatment commenced 7 days after implantation of the tumors, when palpable tumors (mean volume, 65 mm3) were evident in the animals (designated as day 0).
- Fig. 3.
Effect of SAHA treatment on the peripheral blood PSA levels in mice bearing CWR22 prostate xenografts and PSA mRNA expression in the tumors. A, levels of PSA protein in the serum (shown as mean ± SE) were measured in all mice treated daily with vehicle or SAHA (25, 50, or 100 mg/kg) for 21 days. PSA was detected by RIA in serum taken from each mouse by tail-bleed on the first day of treatment (day 0), day 9 of treatment, and at the end of the experimental period (day 21). B, the expression of PSA mRNA was determined by Northern blot using total RNA extracted from two tumors in each treatment group. The top panel shows the PSA mRNA levels in the tumors detected using a 32P-labeled 510-bp cDNA probe corresponding to a region within the open reading frame of the human PSA protein. The ethidium bromide-stained 18S rRNA on the blot is shown in the bottom panel to indicate RNA loading.
- Fig. 4.
Accumulation of acetylated core histones H3 and H4 in CWR22 tumors after administration of SAHA. Mice with palpable CWR22 tumors were given a single i.p. injection of vehicle, 25 mg/kg SAHA, or 50 mg/kg SAHA. Tumors were removed 6 or 12 h after administration of the agent. Two mice were analyzed for each dose at each time point. Acetylation of histones H3 and H4 was analyzed by Western blotting of histone samples (2.5 μg) using antibodies against acetylated histone H3 (top panel) or H4 (bottom panel). The Coomassie blue-stained polyacrylamide gel of histone samples extracted from the tumors is shown beneath each Western blot.
Tables
- Table 1
Effect of SAHA administration on the body weights and survival of the CWR22-bearing mice
Treatment Initial body weight (g)a Final body weight (g) Deaths [days (D) of treatment] Vehicle (DMSO) 26.2 ± 0.5 (n = 8) 27.7 ± 0.6 (n = 8) 0 25 mg/kg SAHA 23.8 ± 0.7 (n = 8) 25.5 ± 1.3 (n = 8) 0 50 mg/kg SAHA 24.8 ± 0.7 (n = 9) 25.8 ± 0.7 (n = 8) 1 (D20) 100 mg/kg SAHA 24.8 ± 0.7 (n = 9) 24.0 ± 0.8 (n = 4) 5 (D6, D9, D14, D16, and D21)b -
a All values are presented as the mean ± SE for the surviving animals (n) in each treatment group. Initial and final body weights refer to the weight of the mice at the start of SAHA treatment and the end of the experiment, respectively.
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b Mice that died in this dose group showed marked weight loss for 1–2 days before death.
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Volume 60, Issue 18
