NSCLC Xenograft Mice Models

Female Balb/c-nude mice (4–6 weeks, 16–20 g) were purchased from Laboratory Animal Center of Sun Yat-sen University [Guangzhou, China; License number: SCXK (Guangdong) 2016-0029], and kept in the animal facility in the SPF animal laboratory [License number: SYXK (GZ) 2014-0144] at International Institute for Translational Chinese Medicine, Guangzhou University of Chinese Medicine (Guangzhou, China). The animal experiments were approved by the Guangzhou University of Chinese Medicine Animal Care and Use Committee (Guangzhou, China), and conducted according with the ethical standards and national guidelines. For A549 xenograft model, mice were subjected to subcutaneously injection with A549 cells (2.5 × 10⁶ cells/mouse) in each right flank. Tumor volume (TV), defined based on two dimensions (L, long diameter; W, wide diameter), was measured by calipers, and calculated as formula: TV (mm³) = (L × W²)/2. When the tumors reached a mean volume of 50 mm³, eligible mice were randomized into four groups (n = 7) as following: gavage control (sterilized coin oil), afatinib (5 mg/kg), SHK (5, 10 mg/kg). For H1299 xenograft model, mice were subjected to subcutaneously injection with H1299 cells (3 × 10⁶ cells/mouse) in each right flank. When the tumors reached a mean volume of 50 mm³, the mice were randomized into two groups (n = 7) as following: gavage control (sterilized coin oil), SHK (10 mg/kg). Animals received oral administration with drugs five times a week for 4 weeks. Body weights were recorded every 3 days. At the end of the experiments, mice were sacrificed, and the tumors and organs were removed and weighed for following assays.