Colon adenocarcinoma HCT116 and HT-29 human colon cancer cells are used. The CellTiter-Glo Luminescent Cell Viability Assay is used to measure cell viability, which use a unique, stable form of luciferase to measure ATP as an indicator of viable cells, and the luminescent signal produced is proportional to the number of viable cells present in culture. Cells are pretreated with Mitomycin C (5 μM) for 12 h or 24 h, and then exposed to different concentrations of TRAIL for 12 h. An equal volume (100 μL) of CellTiter-Glo^TM reagent is added and the solution is mixed gently for 2 min on an orbital shaker. Mixture is incubated at room temperature for 10 min to allow luminescent signal to stabilize, and then imaging is performed using the Xenogen IVIS system to quantify the cell viability^[1].

MCE has not independently confirmed the accuracy of these methods. They are for reference only.

Mice^[1]
Four- to 6-wk-old NCr nude mice are treated with Mitomycin C (1 mg/kg) by intraperitoneal injection for 24 h, followed by one intravenous dose of purified rhTRAIL (100 μg). As a negative control, a subset of the mice are injected (i.p. and i.v.) with saline (vehicle) at the same frequency of treatment. Animals are treated for 3 consecutive weeks. The tumor size is monitored every week using caliper measurements of the tumor volume.
Rats^[3]
Young adult female Wistar rats at age 13 weeks with a median weight of 217 g (range 187 to 255) are randomized into 4 groups of 10 each, namely a normal group with no instillations, an NaCl 0.9% or placebo group that received instillations with the solvent of the chemotherapeutic agents, an Mitomycin C (1 mg/mL) group and an Epirubicin (1 mg/mL) group.

MCE has not independently confirmed the accuracy of these methods. They are for reference only.

• [1]. Cheng H, et al. Mitomycin C potentiates TRAIL-induced apoptosis through p53-independent upregulation of death receptors: Evidence for the role of c-Jun N-terminal kinase activation. Cell Cycle. 2012 Sep 1;11(17): 3312-23.

  [2]. Abbas T, et al. Differential activation of p53 by the various adducts of mitomycin C. J Biol Chem. 2002 Oct 25;277(43):40513-9.

  [3]. Michielsen D, et al. Mitomycin C and epirubicin: functional bladder damage in rats after repeat intravesical instillations. J Urol. 2005 Jun;173(6):2166-70.

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