Table 1

EB1089 induces G0-G1 accumulation and DNA fragmentation

LNCap cells were plated at a density of 250,000 cells/10-cm plate and treated for 6 days with either vehicle (0.1% ethanol), calcitriol, or EB1089. For cell cycle experiments, cells were pulsed before harvesting for 15 h with bromodeoxyuridine. Cells were fixed in ethanol and bromodeoxyuridine incorporation detected using an anti-bromodeoxyuridine antibody. G0G1 represents cells not labeled with antibody. For DNA fragmentation experiments, cells were labeled with terminal transferase following the Boehringer Mannheim protocol. In both experiments, cells were counterstained with propidium iodide, and staining/labeling was quantitated using a flow cytometer. Numbers represent mean cell population ± SD (n = 3). The experiments were performed a minimum of three times, and a representative experiment is shown. ANOVA was performed to determine statistical significance between treatment groups.

Treatment% G0-G1% Td+ labeling
Ethanol22.4 ± 16.83.0 ± 1.6
1 nm calcitriol26.5 ± 1.5
10 nm calcitriol17.1 ± 4.3a
100 nm calcitriol23.2 ± 0.0a
0.1 nm EB108942.1 ± 1.1a
1 nm EB108985.3 ± 1.6b22.9 ± 6.5a
10 nm EB108989.0 ± 1.5a
  • a P ≤ 0.05 compared with ethanol sample.

  • b P ≤ 0.05 compared with same concentration of calcitriol.