Twenty balb c nude mice

Twenty BALB/c nude mice (5-week-old, 14–20 g) supplied by Beijing Vital River Laboratory Animal Technology Co., Ltd. (Beijing, China) were used in this study. For the in vivo experiment, SGC-7901 cells transfected with pLVX-Puro-TRIM50 or vector were subcutaneously implanted to the mice. Then, based on the injected cells, the mice were separated into 2 groups (Vector, cells transfected with vector; TRIM50, cells transfected with pLVX-Puro-TRIM50, n = 6). All animals were housed in a standard environment. Tumor volume was recorded at an interval of 3 days by the formula of 0.5 × width² × length from the 7th day. Thirty-one days after injection of transfected cells, the animals were euthanized, and tumors were resected, weighed, and subjected to Western blot for the detection of TRIM50, β-catenin, Cyclin D1, c-Myc and E-cadherin protein expression. All animal experiments were performed with the approval of the Animal Experimentation Ethics Committee of Zhejiang Eyong Pharmaceutical Research and Development Center (Certificate No. SYXK (Zhe) 2021–0033), and the experiments were conducted strictly following the guidelines of the Institutional Animal Care and Use Committee.

Twenty BALB/c nude mice, 6 weeks old, were provided by Beijing Vital River Laboratory Animal Technology Co. Ltd. All the animals were housed according to a previously described approach (15 (link)). Mice were randomly divided into four groups (n = 5/group): experimental group 1, which was subcutaneously injected with circRNA_103862 downregulated TU212 cells in the dorsal region; experimental group 2, which was subcutaneously injected with circRNA_103862 overexpression AMC-HN-8 cells in the dorsal region; two control groups that were injected with the same amount of TU212 or AMC-HN-8, respectively. The size of the tumor was measured twice a week with calipers, and the volume of xenografts was determined using the formula of a rotational ellipsoid (length × width² × 0.5). Mice were euthanized 3 weeks after cell injection, and the tumor was collected and further analyzed. LSCC cell proliferation was examined using a proliferating cell nuclear antigen (PCNA) in LSCC xenografts.