All animal experiments were approved by the Ethical Committee of Sun Yat-Sen University (approval no. SYSU-IACUC-2020-000562) and were performed in accordance with the guidelines for animal care and protection (27). A total of 14 BALB/c nude mice (female; age, 4-6 weeks old; weight, 18-20 g) were obtained from GemPharmatech Co., Ltd., and were housed in specific pathogen-free conditions at 25°C and 50% humidity, with a 12-h light/dark cycle and free access to food and water. The mice were randomly divided into 2 groups (n=7). A total of 2x106 SCC-15 cells with or without miR-411-5p expression knockdown were suspended in 100 μl PBS, and then injected into the tongue of the nude mice following anesthesia by isoflurane [4% (0.5 l/min) for induction and 2% for maintenance]. Following 8 weeks, the maximum tumor diameter in the mice was recorded as 5.65 mm and the maximum tumor volume was 62.40 mm3, all mice were sacrificed using cervical dislocation and death was confirmed by the absence of breathing, and the lack of a heartbeat and corneal reflex. Finally, the cervical lymph nodes were collected and analyzed using IHC analysis.

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