In vivo tumor xenograft study

Male Balb/c nude mice at the age of 4-6 weeks were obtained from Shanghai SLAC laboratory animal co. Ltd (Shanghai, China). The animal experiments were carried out in accordance with the protocols approved by the Institutional Animal Care and Use Committee of The Affiliated Hospital of Kunming Medical University. SW480 cells were transfected with SPRY4 knockdown plasmid or SPRY4 overexpression plasmid. The transfected cells were subcutaneously injected into the axilla of the mice. GSK126 (an inhibitor of EZH2, Selleck) was administrated intravenously at a dose of 150 mg/kg. The tumor volume was calculated as follows: tumor volume = 1/2 × length ×width². After 28 days, the mice were euthanized, and the tumors were weighed and collected for further tests.