Doxorubicin (Adriamycin) is a Topoisomerase II (Top2) inhibitor with antineoplastic activity.

Topo I:0.8 μM (IC50), Topo II:2.67 μM (IC50)

在高难度测试下（Doxorubicin为2 μM，Simvastatin为10 μM），Doxorubicin与Simvastatin的组合能够杀死97%的Hela细胞[2]。

携带PC3异种移植瘤的小鼠分别被注射2、4或8 mg/kg的Doxorubicin，并随时间测量肿瘤体积。2 mg/kg的剂量对肿瘤生长无影响，而更高剂量初期可以延迟肿瘤生长（第18天和第22天时p<0.05），4 mg/kg或8 mg/kg的Doxorubicin显著降低PC3异种移植瘤中c-FLIP的水平[3]。在大鼠中，分别给予单次腹腔注射10 mg/kg（Doxorubicin 1）、连续10天每天腹腔注射1 mg/kg（Doxorubicin 2）或每周腹腔注射2 mg/kg共5次（Doxorubicin 3）。在Doxorubicin 1处理组中，第28天观察到80%的死亡率，而Doxorubicin 2和Doxorubicin 3分别在第107天和第98天达到80%死亡率。在第2周时，Doxorubicin 1导致的分数缩短降低了30%，在第13周Doxorubicin 2和Doxorubicin 3分别降低了55%和42%[4]。

Doxorubicin is dissolved in stock solutions (1 mM) and serially diluted with RPMI 1640 media (0.1, 1, and 2 μM)[2]. 160 μL of Hela cells suspension (3×104 cell/mL) is dispensed into three 96-well U-bottom microplates and incubated for 24 h at 37°C in a fully humidified atmosphere of 5% CO2. In plate 1, serial dilutions of Doxorubicin (20 μL; final concentration, 0.1-2 μM) and Simvastatin (20 μL; final concentration, 0.25-2 μM) are added to a final volume of 200 μL and incubated for another 72 h. In plates 2 and 3 serial dilutions of each drug (Simvastatin or Doxorubicin, 40 μL) are added. After an incubation period of 24 h, the medium is aspirated and the cells are washed in PBS. Then, serial dilutions of other drug (40 μL) are added and supplemented with culture medium to a final volume of 200 μL, and incubated for 48 h. Doxorubicin and Simvastatin are used individually as positive controls (40 μL in each well), and the cells treated only with solvent are considered as negative controls. To evaluate cell survival, 20 μL of MTT solution (5 mg/mL in PBS) is added to each well and incubated for 3 h. Then the media is replaced with 150 μL of DMSO, and complete solubilization of formazan crystals is achieved by repeated pipetting of the solution. Absorbance is then determined at 540 nm by an ELISA plate reader. Each drug concentration is assayed in 4 or 8 wells and repeated 3 times. The cytotoxic/cytostatic effect of Doxorubicin is expressed as the relative viability (% control) and calculated. Percentage of cell survival in the negative control is assumed as 100. Relative viability=(experimental absorbance-background absorbance)/ (absorbance of untreated controls-background absorbance)×100 %[2].