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Research Area
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Description
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Sarcoma,Medulloblastoma, Medulloblastoma,Cancer |
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Protocol
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Cell Assay
[2]
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| Cell Lines |
MDCKII cells |
| Concentrations |
20 μM |
| Incubation Time |
2 hours |
| Methods |
MDCKII cells are seeded into 24-well plates at a density of 3 × 105 cells per well and are allowed to attach. Medium is then changed to that containing different drugs (50 μM VP, 50 μM indomethacin, or 20 μM GDC-0449 in DMSO or DMSO alone as control, and nonfluorescent calcein-AM is added to a final concentration of 1.0 μM and incubated at 37 °C for 2 hours. Cells are then washed twice with Ca2+, Mg2+-containing Hank's balanced salt solution buffer and lysed by shaking in 0.01% Triton X-100 in PBS buffer for 1 hour at room temperature or overnight at 4 °C. The lysate is then transferred into 96-well plates, and the fluorescence signal caused by the cell-derived calcein was quantified spectrophotometrically with a SpectraMax M5 Multi-Detection Readerusing an excitation wavelength of 495 nm and an emission wavelength of 515 nm. All manipulations are performed in the dark. All readings are expressed as mean ± SEM normalized to the control. |
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Animal Study
[4]
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| Animal Models |
Ptch(+/-) allograft model, D5123 and 1040830 |
| Formulation |
In 0.5% methyl-cellulose, 0.2% tween-80 |
| Doses |
~ 100 mg/kg |
| Administration |
Orally |
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References |
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[1] Scales SJ, et al. Trends Pharmacol Sci. 2009, 30(6), 303-312.
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[2] Zhang Y, et al. Neoplasia. 2009, 11(1), 96-101.
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[3] Tian F, et al. Anticancer Res. 2012, 32(1), 89-94.
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[4] Wong H, et al. Clin Cancer Res. 2011, 17(14), 4682-4692.
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