Tumor xenograft studies in nude mice

Taxol resistant NSCLC cells (TR A549 cells) were used to determine the in vivo efficacy of WFA. Female athymic nude (nu/nu) mice (5–6-week old) were purchased from Charles River Labs and maintained in accordance with the Institutional Animal Care and Use Committee guidelines. TR A549 cells (2.5 × 10⁶ cells/mouse) in 100 μL of serum-free media were mixed (50:50) with Matrigel matrix (Becton Dickinson, Bedford, MA) and subcutaneously injected into the left flank of each mouse. The mice were provided purified AIN-93M diet and water ad libitum. Once average tumor size reached about 80–120 mm³, mice were randomly divided into 3 groups (n = 5): (i) treated with vehicle, (ii) PAC (10 mg/kg divided in 3 doses per week) and (iii) WFA (10 mg/kg) as 3 doses per week. All treatments for vehicle and intervention drugs were done via intraperitoneal injections (i. p) and the tumor volumes were measured twice weekly.