Article Figures & Data
Figures
- Figure 1.
In vitro reduction of NICD, Hes-1, and cMyc by PF-03084014 in HPB-ALL Cells. A, lysates of HPB-ALL cells treated with PF-03084014 at indicated concentrations for 72 hours were subjected to immunoblot analysis for NICD. B, HES-1 and cMyc expression was quantified by real-time PCR analysis. Student's t test was used to determine the P value (*, P < 0.05; **, P < 0.01).
- Figure 2.
Induction of cell cycle arrest and apoptosis by PF-03084014 in T-ALL cells. A, HPB-ALL cells were treated with indicated concentrations of PF-03084014 for 7 days. G0-G1 cell cycle block was evaluated by propidium iodide (PI) staining of DNA content of the cells. The percentages of cells in each phase are indicated in the graphs. B, cell lysates from A were subjected to immunoblot analysis for NICD, total Rb, P27 Kip1, and actin. C, HPB-ALL and TALL-1 cells were treated with PF-03084014 at indicated concentrations for 7 days. Caspase-3 activities were measured. Results represent the average + SE from triplicates. RLU, relative luciferase activity; EC50, half maximal effective concentration. D, HPB-ALL cells were treated with 1 μmol/L PF-03084014 for indicated time. Cell lysates were subjected to immunoblot analysis for NICD, caspase-3, cleaved PARP, and actin.
- Figure 3.
Time course for induction of cell cycle arrest and apoptosis by PF-03084014. HPB-ALL cells were treated with 1 μmol/L PF-03084014 for indicated time. On day 7 after treatment, media containing compound were removed and cells were incubated in regular growth media for additional 1, 3, and 7 days. A, G0-G1 cell cycle block was evaluated by PI staining of DNA content of the cells. B, apoptosis was measured by FITC-labeled Annexin V. C, cell lysates were subjected to immunoblot analysis for NICD, total Rb, P27 Kip1, cleaved PARP, and actin.
- Figure 4.
Reduction of NICD and tumor growth inhibition by PF-03084014 in HPB-ALL xenograft model. Athymic mice bearing HPB-ALL tumors were orally given PF-03084014 at the indicated dose or vehicle alone over the designated treatment schedule. A, plasma and tumor samples were harvested at designated time points post single dose at 50 mg/kg. Tumor lysates were subjected to immunoblot analysis for NICD. Data were presented as % NICD inhibition (inh.) of treatment versus vehicle (veh.) groups. Plasma samples were analyzed for PF-03084014 concentrations using liquid chromatography/mass spectrometry analysis. B, PF-03084014 was administered twice daily (bid) for indicated time. Tumor volume was measured using Vernier calipers on the indicated days with the median tumor volume ± SE indicated for groups of 10 to 15 mice. C, tumors from mice receiving 150 mg/kg PF-03084014 twice daily for 7 days were resected and fixed in 10% neutral buffered formalin. Fixed tumors were cut into 4-mm sections, and immunostained for NICD (red arrows), cleaved caspase-3 (red arrows), or Ki67. Photomicrographs were taken at 20× (NICD), 10× (caspase-3), and 40× (Ki67) magnifications.
- Figure 5.
Intermittent dosing (7-day on/7-day off) of PF-03084014 reduces its gastrointestinal toxicity and is efficacious in tumor growth inhibition in HPB-ALL model. PF-03084014 was administered twice daily(bid) for indicated time. A, body weight was monitored and recorded. Percentage of change in body weight was presented. B, tumor volume was determined as in Fig. 4.
- Figure 6.
Dexamethasone (Dex) reduces GSI-induced goblet cell hyperplasia in mice. A, histologic analysis of small intestine from mice continuously treated with vehicle (14 d), PF-03084014 (14 d; 150 mg/kg, orally), Dex (14 d; 15 mg/kg, i.p.), or Dex (14 d) plus PF-03084014 (14 d). B, histologic analysis of small intestine from mice continuously treated with vehicle (14 d), PF-03084014 (14 d), PF-03084014 (14 d) plus Dex (7 d on/7 d off), or PF-03084014 (14 d) plus Dex (7 d off/7 d on). Red arrows indicate examples of goblet cell hyperplasia.
Additional Files
Supplementary Data, Wei, et al.
Files in this Data Supplement:
Volume 9, Issue 6
