IMD-0560

Cell experiment:

Cell migration is assessed in a modified chamber containing a gelatin-coated porous membrane. IMD-0560 is placed in the upper chamber 2 hrs prior to TNFα treatment, and TNFα is placed in the lower chamber at 10 ng/mL; then, the chamber is incubated for 24 hrs. Next, the cells attached to the upper surface of the membrane are scraped off, and the cells that migrate to the lower surface are fixed and stained with DAPI[1].

Animal experiment:

Sixty male mice, weighing approximately 20 g at 8 to 10 weeks of age are used. They are anesthetized using ether, and 0.1 mL of SCCVII in DMEM is injected into the left masseter region. The mice are randomly separated into six groups with similar average body weights. One week (early treatment: E) or 2 weeks (late treatment: L) after injection, the mice are injected between the left masseter region and the surface of the left mandibular bone with vehicle (50 μL of carboxymethyl-cellulose: CMC/mouse, n=10) or IMD-0560 (3 or 5 mg/kg/50 μL of CMC/mouse, n=10), 3 times per week for a total of 3 weeks (E) or 2 weeks (L). The tumor sizes are assessed using calipers, and the tumor volume is calculated. At the end of week 3, all surviving mice are euthanized, and the heads of the mice are fixed in 3.7% formaldehyde[1].

References:

[1]. Tada Y, et al. The novel IκB kinase β inhibitor IMD-0560 prevents bone invasion by oral squamous cell carcinoma. Oncotarget. 2014 Dec 15;5(23):12317-30.