TBB

Cell lines

PC-3 cells

Preparation Method

PC-3 cells were cultured routinely in RPMI-1640 media, respectively, which were supplemented with 10% FBS, penicillin (100U/ml) and streptomycin (100µg/ ml) at 37˚C in a humidified atmosphere of 5% CO₂. Cells were seeded at 5x10⁴cells/well in 24-wells plates and cultured for 72h. TBB (final concentration 60µM), Camptothecin (CPT) (final concentration 5.8nM), 2-deoxyglucose (2-DG; final concentration 0.5mM) or tumor necrosis factor-related apoptosis-inducing ligand (TRAIL) (final concentration 13.3ng/ml) were added to the medium individually or in a combination for 24h. Then the medium with the agent was removed and the cells were cultured for another 24h in normal medium. After that, 500µl of MTT mixture (0.5mg/ml in medium without phenol red) was added to each well and incubated for an additional 1h at 37˚C. The formazan crystals were diluted in 250µl of DMSO. The absorbance was measured at 570nm.

Reaction Conditions

60μM; 24h

Applications

TBB reduced the viability of PC-3 cells either alone or combined with anticancer agents CPT or TRAIL when a proper time schedule of the administration is applied.

Animal models

C57BL/6J mice

Preparation Method

Proliferative retinopathy in neonatal mice was induced in heterozygous C57BL/6J mice. Mice from the retinopathy group were placed in 75% oxygen at postnatal day 7 and maintained in these conditions with their nursing mothers for 5 days. These mice were then returned to normal air and maintained for another 5 days. Nor-moxic control mice were maintained in normal air for the same duration as test mice and under the same conditions of light cycle and temperature. For in vivo administration, the CK2 inhibitors emodin and TBB were dissolved in PBS (pH 7.2) with 20% polyethylene glycol 400 (PEG 400) and 2% Tween-80. This solvent did not elicit any animal tolerance problems, unlike ethanol or DMSO. Inhibitors were injected intraperitoneally in volumes of 50μL or less per mouse at doses of 15 to 30mg/kg body weight, twice daily from day 11 to day 17. Control mice were injected with PEG-Tween vehicle alone. At the end of the experiment, mice were euthanatized by intraperitoneal injection with tribromoethanol (0.1ml/g body weight).The eyes were enucleated and fixed in 4% paraformaldehyde in 0.1M phosphate buffer (pH 7.4) for 18 hours for further analysis.

Dosage form

30 to 60mg/kg/day for 6 days; i.p.

Applications

TBB reduced retinal neovascularization by approximately 60% in the mouse OIR model with minimal effects on the main vascular tree but marked suppression or elimination of neovascular tufts.

References:
[1] Orzechowska, E., Koz?owska, E., Staro?, K., & Trzci?ska-Danielewicz, J. (2012). Time schedule-dependent effect of the CK2 inhibitor TBB on PC-3 human prostate cancer cell viability. Oncology reports, 27(1), 281–285.
[2] Ljubimov, A. V., Caballero, S., Aoki, A. M., Pinna, L. A., Grant, M. B., & Castellon, R. (2004). Involvement of protein kinase CK2 in angiogenesis and retinal neovascularization. Investigative ophthalmology & visual science, 45(12), 4583–4591.