Establishment of thyroid tumor xenograft and treatment

The mice were randomly assigned into 3 groups of 10 each: normal control, untreated, and JR-P(II) treatment. The mice in the untreated and JR-P(II) treatment groups were intraperitoneally injected with ketamine hydrochloride plus xylazine hydrochloride anesthesia. SW1736 cells (2×10⁶) in 100 μL PBS were injected subcutaneously into the flanks of mice. Mice in the treatment group were intraperitoneally injected with 24 mg/kg of JR-P(II) daily for 14 days. Mice in the normal and untreated groups received equal volumes of normal saline. Changes in body weight were measured on alternate days and tumor volumes were recorded using electronic calipers. Five mice from each group were sacrificed using carbon dioxide on day 7 and 5 on day 14 of treatment to excise the tumors. The tumors were treated with protein extraction buffer, homogenized, and sonicated on ice. The tissue lysate was centrifuged to collect the supernatants, which were stored in liquid nitrogen until further analysis. The expression of proteins was measured by Western blotting. Body weights of mice were recorded on alternate days during 14 days (on days 2, 4, 6, 8, 10, 12, and 14 of the treatment), and the tumor weights were measured on days 7 and 14.