A total of 12 female athymic BALB/c nude mice (age, 4-5 weeks; weight, 16-18 g) were obtained from the Model Animal Research Center of Soochow University (Suzhou, China). The mice were housed in specific pathogen-free barrier facilities in a temperature-controlled room (24°C) with a 12 h light/dark cycle and 35-40% relative humidity, and were permitted free access to food and drinking water. The body weight of each mouse was measured every 2 days. The mice were humanely sacrificed via cervical dislocation at the study endpoint. All experiments were performed in accordance with the Guide for the Care and Use of Laboratory Animals (NIH publication 80-23, revised 1996) (20), with the approval of the Ethics Committee of Soochow University (approval no. SUDA20201027A01).

To obtain stable cell lines, lentivirus supernatant of sh-GMPS and sh-control (GeneChem, Inc.) were added to HeLa cells at MOI 10, according to the manufacturer's instructions, and incubated at 37°C and 5% CO2 for 48 h, followed by screening with 4 μg/ml puromycin for 2 weeks. Control HeLa cells and GMPS-knockdown HeLa cells (2×106 cells) were suspended in 100 μl PBS media and were subcutaneously inoculated into the flanks of the mice. There were six mice in each group (control and sh-GMPS). Mice were monitored every 2 days for tumor growth, and tumor size was measured using a caliper. The tumor volume was calculated using the modified ellipsoid formula: Volume=1/2 × (length × width2). After 5 weeks, the mice were sacrificed and tumor weight was measured. The maximum volume of the tumors was 1.17 cm3, and the maximum diameter of the tumors was 1.21 cm. A section of each tumor tissue was fixed in 10% formalin at room temperature for 24 h, and used for IHC. The remaining tumor tissues were used for protein and mRNA expression analyses, and the experiment methods were the same as above.

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