Phosphate buffered saline (pbs)

1D-NMR experiments including ¹H NMR (500 MHz) and APT ¹³C NMR (125 MHz) and 2D-NMR experiments including ¹H−¹H COSY, HSQC, HMBC, and NOESY were recorded on a Bruker Avance III HD 500 NMR instrument (Karlsruhe, Germany). Chromatographic techniques were applied to separate compounds, including open columns chromatography using silica gel 60 (0.063 - 0.200 mm), flash column chromatography (FCC) using silica gel 60 (0.040 - 0.063 mm), and thin layer chromatography (TLC) using pre-coated silica gel 60 F254 plates (Merck, Darmstadt, Germany). The solvents were obtained from Merck (Darmstadt, Germany). A human breast adenocarcinoma (MCF-7) cell line was obtained from the Iranian Biological Resource Center, Tehran, Iran. RPMI 1640, phosphate-buffered saline (PBS), and fetal bovine serum (FBS) were acquired from Biosera (Ringmer, UK). Penicillin/streptomycin and cisplatin were obtained from EBEWE Pharma (Unterach, Austria). Also, 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) was taken from Sigma-Aldrich (St. Louis, MO, USA). Hydrochloric acid 32% and chloramphenicol were acquired from Sigma-Aldrich. Finally, p-iodonitrotetrazolium violet (INT) was obtained from Fluka (Germany).

Twenty-five healthy male athymic BALB/c (nu/nu) nude mice were purchased from the SLAC Laboratory Animal Co., Ltd. (Shanghai, China). The mice were 6 weeks old and weighed 18–20 g. The mice were housed and fed in a specific pathogen-free (SPF) environment and provided with sterile food and water. All procedures were performed under aseptic condition. Before the animal experiments, the mice were fed ad libitum for one week in the new environment.
Cultured cells were washed with phosphate-buffered saline (PBS) (Biosera, Boussens, France), and 0.2 mL of cell suspension (1×10⁷ cells) was injected into the skin of the right underarm region of five nude mice.