Establishment of 4T1 Breast Cancer Mouse Model and Animal Administration

4T1 cells were cultured in large quantities and digested when they reached the logarithmic growth phase. BALB/c mice were transplanted subcutaneously into the right axilla with 0.1 mL of cell suspension containing 1×106 4T1 cells to establish the subcutaneous xenograft mouse model of breast cancer. The weight of mice and the growth of tumors were measured daily. When the tumors grew to 100 mm3 in size, the mice were randomly divided into five groups: the DMDD-H group (DMDD 100 mg/kg, qd, ig), the DMDD-M group (DMDD 50 mg/kg, qd, ig), the DMDD-L group (DMDD 25 mg/kg, qd, ig), the DOX group (doxorubicin 0.5 mg/kg, qw, ip) and the model group, with 10 mice in each group. In addition, 10 mice without tumor cells were selected as the normal group. Mice were treated for 14 days, with tumor volume and weight were measured every two days. Tumor volume was calculated as (width2 × length)/2. The inhibition rate of tumor growth was calculated as (%)= (Mean tumor weight of vehicle group − Mean tumor weight of treatment group)/Mean tumor weight of vehicle group × 100%. In addition, the liver index = liver weight (mg)/body weight (g), and the spleen index = spleen weight (mg)/body weight (g).