Female balb c athymic nude mice

Female BALB/c athymic nude mice (5 weeks) with body weight from 18 to 22 g were purchased from the Model Animal Research Center of Nanjing University. About 3 × 10⁶ NCI-H460 cells were injected into the subcutaneous tissue of armpit. Tumor tissues were grown with a volume about 400 mm³, then resected and cut into small pieces. Subsequently, the pieces of tissue were planted subcutaneously into each nude mice. After 10 days, tumor sizes were measured by micrometer calipers. After excluding the mice with unsuitable tumor size, the mice with analogous TV were randomly divided into five groups with six individuals per group. DT-13 was intragastrically administrated with a concentration of 1.25 mg/kg, and NVB was intravenously administrated with dosages of 1 and 10 mg/kg (positive control). The negative group was given an equal amount of normal saline. After administration for 21 days, mice were killed, and the tumor tissues were then resected and detected. TV and RTV were calculated by the following formula: TV (mm³)=A/2 × B², where A represents the longest diameter of tumor, and B represents the shortest diameter. RTV=V_t/V₀, where V_t represents the TV of day t, and V₀ represents the TV of day 0. Animal care and surgery operation were all guided by Animal Care and Control Committee in China Pharmaceutical University.

Female BALB/c athymic nude mice (5 weeks) with body weight from 18 to 22 g were purchased from the Model Animal Research Center of Nanjing University. Mice were maintained according to the guidelines for the welfare and use of animals in cancer research in a temperature-controlled room (22°C). THP-1 macrophages were firstly activated by 15 mM lactic acid, and then co-cultured with MDA-MB-231 cells for seven days. Culture media were changed every three days. After co-culture, breast cancer cells were collected and 3×10⁵ MDA-MB-231 cells were injected into the tail vein of nude mice. After two weeks, animals were sacrificed and metastatic nodules on lung surfaces were counted. To validate the effects of CCL5 in promoting lung metastasis, 5μg/ml anti-CCL5 neutralizing antibody was added to the co-culture system.

Female BALB/c athymic nude mice (5–6 weeks) with body weights from 18 to 22 g were purchased from the Model Animal Research Center of Nanjing University (Nanjing, China). A total of 2 × 10⁶ MDA-MB-231 cells transfected with shControl or shSLC1A5#1 were injected into the subcutaneous tissue of the armpit. Tumours were grown until their volume reached 300 to 500 mm³, resected, and cut into small pieces. Subsequently, the tissue pieces were subcutaneously implanted into each of the nude mice. The mice were randomly divided into groups of six individuals each. C118P was administered by tail vein injection at a concentration of 50 mg/kg. The negative group was given an equal amount of normal saline. At 21 days after administration, the mice were euthanised, and the tumour tissues were then resected and assessed. Tumour volume (TV) was calculated by the following formula: TV (mm³) = A/2 × B², where A represents the longest diameter of the tumour, and B represents the shortest diameter. Relative tumour volume (RTV) was calculated with the following formula: RTV = V_t/V₀, where V_t represents the TV on day t, and V₀ represents the TV on day 0. The animal care and surgical procedures were guided by the Animal Care and Control Committee of China Pharmaceutical University.