|
Research Area
|
|
Description
|
Solid tumours, Non-small cell lung cancer |
|
Protocol
|
|
Kinase Assay
[1]
|
| Kinetic Methods |
In a typical kinetic run, 2.00 mL of assay buffer (20 mM HEPES, 0.5 mM EDTA, 0.035% SDS, pH 7.8) and Suc-Leu-Leu-Val-Tyr-AMC in DMSO are added to a 3 mL fluorescence cuvette, and the cuvette is placed in the jacketed cell holder of a fluorescence spectrophotometer. Reaction temperature is maintained at 37 °C by a circulating water bath. After the reaction solution has reached thermal equilibrium (5 minutes), 1 μL?10 μL of the stock enzyme solution is added to the cuvette. Reaction progress is monitored by the increase in fluorescence emission at 440 nm (λex = 380 nm) that accompanies cleavage of AMC from peptide-AMC substrates. |
|
Cell Assay
[1]
|
| Cell Lines |
U266, IM-9, and Hs Sultan cells |
| Concentrations |
0-10 μM |
| Incubation Time |
48 hours |
| Methods |
The inhibitory effect of Bortezomib on MM and BMSC growth is assessed by measuring MTT dye absorbance of the cells. Cells from 48-hour cultures are pulsed with 10 μL of 5 mg/mL MTT to each well for the last 4 hour of 48-hour cultures, followed by 100 μL of isopropanol containing 0.04 N HCl. Absorbance is measured at 570 nm using a spectrophotometer. |
|
Animal Study
[2]
|
| Animal Models |
Mice are inoculated s.c. into the right flank with RPMI 8226 MM cells. |
| Formulation |
Bortezomib is dissolved in 0.9% sodium chloride. |
| Doses |
≤1mg/kg |
| Administration |
Administered via i.v. |
|
References |
|
[1] Hideshima T, et al. Cancer Res, 2001, 61(7), 3071-3076.
|
|
[2] LeBlanc R, et al. Cancer Res, 2002, 62(17), 4996-5000.
|
|
[3] Yu C, et al. Blood, 2003, 102(10), 3765-3774.
|
|
[4] Nie D, et al. Leuk Lymphoma, 2012.
|
|