HCC Xenograft Animal Model Study

Murine experiments were approved by the University of Pittsburgh Institutional Animal Care and Use Committee and the Second Hospital of Anhui Medical University. Transfected Huh7 cells (1 × 10⁷/mice) were injected into the right flank subcutaneously with 200 μl serum-free DMEM in 8-week-old male BALB/c (nu/nu) mice (Jackson laboratory, Bar Harbor, ME). Doxorubicin (4 mg/kg) (Sigma-Aldrich, St. Louis, MO) was injected through tail vein once every 2 days starting 6 days after tumor cell injection. Tumor size and mice weight (volume = length × width × width/2) were measured every 5 days until the 25th day after tumor cell injection. Mice were sacrificed and tumor mass was harvested for immunohistochemistry.