Chlorambucil

Cell experiment:

Cultured cells at log-growth phase are digested by trypsin into single cell suspension and are seeded into 96-well plate at 1000 per well density. The plate is placed in a 37°C chamber with 5% CO2. After attached growth for 24 h, cells are treated with TRAIL at 0, 20, 40 and 80 ng/mL or Chlorambucil at 0, 2.5, 5 and 10 μM for 48 h. 10 μL CCK-8 reagent is added to each well, followed by incubation at 37°C for 4 h. Absorbance values at 450 nm are then measured by a micro-plate reader. Six parallel samples are performed in each treatment group. Cell proliferation rate (%) = mean value of experimental group/mean value of control group × 100%[1].

Animal experiment:

Mice[2]Female Swiss mice are divided randomly into five group (20 mice per group). Group 1 is kept as the control group, Group 2 receives intraperitoneal injection of by 2.5 × 106 Ehrlich ascites carcinoma cell, Gropu 3 is treated orally with Chlorambucil 0.2 mg/kg body weight, Group 4 is treated orally with levamisole (5 mg/kg body weight) and Group 5 is treated orally with a combination of Chlorambucil and levamisole each day, using a bent stainless steel stomach tube[2].

References:

[1]. Guo JX, et al. Synergistic effects of chlorambucil and TRAIL on apoptosis and proliferation of Raji cells. Eur Rev Med Pharmacol Sci. 2017 Oct;21(20):4703-4710.
[2]. Salem FS, et al. Biochemical and pathological studies on the effects of levamisole and chlorambucil on Ehrlich ascites carcinoma-bearing mice. Vet Ital. 2011 Jan-Mar;47(1):89-95.