Mitoxantrone (mitozantrone) is an inhibitor of topoisomerase II (Topo II), an inhibitor of protein kinase C (PKC) (IC50=8.5 μM). Mitoxantrone has antitumor activity for the treatment of acute myeloid leukemia, hepatocellular carcinoma, and breast cancer.

PKC:8.5 μM.

方法：人乳腺癌细胞 MCF7 用 Mitoxantrone (0.07-9.36 µM) 处理 48 h，使用 SRB assay 测定细胞活力。
结果：MCF7 细胞在 Mitoxantrone 1.17 µM 剂量下达到对应于 IC50 的抑制作用。[1]
方法：骨肉瘤细胞 U2OS 用 Mitoxantrone (0.2-1 µM) 处理 48 h，使用 Western Blot 检测靶点蛋白表达水平。
结果：Mitoxantrone 处理增加了 Caspase-3 和 PARP1 的裂解。Mitoxantrone 处理还增加了骨肉瘤细胞中 Bax 和 Bim EL 的表达。然而，Mitoxantrone 处理降低了 Bcl-2 的表达。[2]

方法：为测试对，将 Mitoxantrone (0.4-3.2 mg/kg) 腹腔注射给注射白血病细胞 L1210 的 CDF1 小鼠，每天一次。
结果：以最佳剂量 (1.6 mg/kg/天) 给予 Mitoxantrone 产生了具有统计学意义的 60 天存活率。[3]

activity-based protein profiling (ABPP): Mouse brains are Dounce-homogenized in PBS, pH7.5, followed by a low-speed spin (1,400×, 5 min) to remove debris. The supernatant is then subjected to centrifugation (64,000×, 45 min) to provide the cytosolic fraction in the supernatant and the membrane fraction as a pellet. The pellet is washed and resuspended in PBS buffer by sonication. Total protein concentration in each fraction is determined using a protein assay kit. Samples are stored at -80 °C until use. Mouse brain membrane proteomes, are diluted to 1 mg/mL in PBS and pre-incubated with varying concentrations of inhibitors (1 nM to 10 mM) for 30 min at 37 °C before the addition of FP-rhodamine at a final concentration of 2 mM in a 50 mL total reaction volume. After 30 min at 25 °C, the reactions are quenched with 4×SDS-PAGE loading buffer, boiled for 5 min at 90 °C, subjected to SDS-PAGE and visualized in-gel using a flatbed fluorescence s

The human breast carcinoma cell lines MDA-MB-231 and MCF-7 are seeded in standard 96-well plates. One day after seeding, the culture medium is changed and replaced by medium containing different concentration of Mitoxantrone (10-5 to 5 μM) with or without DHA (30 μM) during 7 days. Viability of cells are measured as a whole by the tetrazolium salt assay[3].