In vivo experiments

A nude mouse xenograft model was established using 4–6 week-old female BALB/c nude mice. Mice were kept in laminar-flow cabinets under specific pathogen-free conditions, and handled according to the recommendations of the National Institutes of Health guidelines for care and use of laboratory animals. 5 × 10⁶ SMMC-7721 cells were inoculated subcutaneously into the flank of each nude mouse. The tumor volume for each mouse was determined by measuring two of its dimensions and then calculated as tumor volume = length × width × width/2. The developing tumors were observed over the next 3 weeks, and the mice were then sacrificed at the end of follow-up. All in vivo experiments protocols were approved by the Institutional Animal Care and Use Committee of Southern Medical University.