Doxorubicin hydrochloride (Adriamycin) belongs to the anthracycline class of antibiotics and is an inhibitor of human DNA topoisomerase I/II (IC50=0.8/2.67 μM). Doxorubicin hydrochloride exhibits cytotoxicity and antitumor activity. Doxorubicin hydrochloride reduces the phosphorylation of AMPK and its downstream target protein acetyl coenzyme A carboxylase, and induces apoptosis and autophagy.

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

方法：人乳腺癌细胞 MCF10A、BT474、MCF-7 和 T47D 用 Doxorubicin hydrochloride (0.1-10 μM) 处理 48 h，使用 MTT 方法检测细胞生长抑制情况。
结果：Doxorubicin hydrochloride 剂量依赖性地抑制 MCF10A、BT474、MCF-7 和 T47D 细胞生长，IC50 分别为 2.51 µM、1.14 µM、0.69 µM 和 8.53 µM。[1]
方法：牛主动脉内皮细胞 BAECs 和 人卵巢畸胎瘤细胞 PA-1 用 Doxorubicin (0.5 μM) 处理 1-16 h，使用 Flow Cytometry 方法检测细胞凋亡情况，使用 caspase-3 assay kit 检测 caspase-3 的活性。
结果：Doxorubicin 时间依赖性地诱导 BAECs 和 PA-1 细胞的凋亡及 caspase-3 激活。[2]
方法：犬乳腺癌细胞 CIPp 用 Doxorubicin (EC50(20h)=12.08 μM) 处理 3-48 h，使用 qRT-PCR 方法检测靶基因表达情况。
结果：Doxorubicin 诱导多药耐药性 (MDR) 相关基因 P-gp 和 BCRP 的 mRNA 表达水平上调。[3]

方法：为检测体内抗肿瘤活性，将 Doxorubicin hydrochloride (1 mg/kg/4 天) 和 lovastatin (5  mg/kg/天) 腹腔注射给携带鼠黑色素瘤肿瘤 B16F10 的 B6D2F1 小鼠，持续两周。
结果：与单独作用的任一药物相比，Doxorubicin hydrochloride 和 lovastatin 联合治疗的敏感性显著增加，lovastatin 增强了 Doxorubicin hydrochloride 的抗肿瘤活性。[4]
方法：为研究 Doxorubicin 对癌症患者的急性和长期认知障碍，将 Doxorubicin hydrochloride (25 mg/kg) 单剂量腹腔注射给 B6C3F1J 小鼠。
结果：Doxorubicin hydrochloride 全身治疗在 24 h 内改变了与认知功能相关的关键细胞核中的谷氨酸神经传递，对空间学习和记忆没有持久影响。[5]

To analyze the effect of Bcl-2 expression on the viability of HUVECs treated with Dox, cells were co-transfected with 200 ng of the pEGFP-spectrin expression plasmid together with 200 ng of either pCDNA3-hBcl-2 or the control pCMVβ-galactosidase expression vector (33). The pGL3 Basic vector (2.1 μg) was added as a DNA carrier in a total volume of 0.140 ml, and transfection was performed by the calcium phosphate procedure in 35-mm tissue culture dishes. After treatment, the cells were washed with PBS, fixed with 3.7% formaldehyde for 15 min, and washed for a further 10 min with 50 mM NH4Cl blocking solution in PBS. Cells were then washed with PBS, permeabilized with a 0.1% Triton X-100 for 10 min, washed again with PBS, and stained with 1 μg/ml 4′,6-diamidino-2-phenyl-indole solution for 2 min. The cells were examined under a fluorescence microscope, and GFP-positive cells were scored after counting a minimum of 1000 total cells for each condition. The efficiency of transfection in Bcl-2- and β-galactosidase-expressing cells, determined in aliquots of transfected cells just before the addition of Dox, was similar (10–12%) [1].

Athymic male nude mice (3-4 weeks old) are used. PC3 cells (4×106) are injected subcutaneously into the flanks of mice. Animals bearing tumors are randomly assigned to treatment groups (five or six mice per group) and treatment initiated when xenografts reached volumes of about 100 mm3. Tumors are measured using digital calipers and volume calculated using the formula: Volume=Width2×Length×0.52, where width represents the shorter dimension of the tumor. Treatments are administered as indicated using vehicle (PBS containing 0.1% BSA), Doxorubicin (2-8 mg/kg), Apo2L/TRAIL (500 μg/animal), or a combination of 4 mg/kg Doxorubicin followed by 500 μg Apo2L/TRAIL. Doxorubicin is administered systemically whereas Apo2L/TRAIL is given either intratumorally or systemically. All treatments are given once. Mice are monitored daily for signs of adverse effects (listlessness and scruffy appearance). Treatments seemed to be well tolerated. The mean±SEM is calculated for each data point. Differences between treatment groups are analyzed by the student t-test. Differences are considered significant when P<0.05 [3]. Altogether, 29 male Wistar rats (weight 306 ± 18.6 g) were used in the study. Animals were divided into three groups: control (group C; n = 10; 306.4 ± 17.2 g), animals treated with DOX (group DOX; n = 10; 305.0 ± 24.9 g) and animals treated with L-DOX (group L-DOX; n = 9; 306.7 ± 15.0 g). Vehiculum (aqua pro injection), DOX and L-DOX were applied to group C, DOX and L-DOX, respectively, by single intraperitoneal injection; concentration of both DOX and L-DOX was 5 mg/kg, similar to the concentrations used in human treatment protocols. All animals were sacrificed 24 h after drug application. Thoracotomy was performed, hearts were excised and samples were obtained separately from the free wall of the left atrium (LA), left ventricle (LV), right atrium (RA) and right ventricle (RV).Samples were placed into RNA later preservation solution and stored at -80 C until further analysis [4].