Docetaxel (RP-56976) is a semi-synthetic analog of paclitaxel, a microtubule depolymerization inhibitor (IC50=0.2 μM). Docetaxel attenuates the effects of bcl-2 and bcl-xL gene expression and exhibits apoptosis-inducing, anti-tumor activity.

Microtubule:0.2 μM

方法：人肺癌细胞 NCI-H460 用 Docetaxel (0.2-200 nmol/L) 处理 24-72 h，使用 MTS 方法检测细胞活力。
结果：NCI-H460 在 72 h 时对 Docetaxel 的 IC50 为 0.030 μmol/L，24 h 时为 0.116 μmol/L。[1]
方法：人前列腺癌细胞 PC-3、DU-145 和 LNCaP 用 Docetaxel (0.5-4 nM) 处理 48 h，使用 Flow Cytometry 检测细胞凋亡情况。
结果：高剂量 Docetaxel 处理显著增加了 Annexin V+ 凋亡细胞的比例。[2]

方法：为检测体内抗肿瘤活性，将 Docetaxel (5-10 mg/kg) 和 PD-1 inhibitor (200 μg/只) 腹腔注射给携带小鼠前列腺癌肿瘤 RM-1 的 CB17 SCID 小鼠，每周五次，持续十天。
结果：PD-1 inhibitor 联合 Docetaxel 对小鼠前列腺癌具有协同作用，抑制了前列腺肿瘤的生长，提高了存活率并减少了不良反应。[3]
方法：为检测体内抗肿瘤活性，将 Docetaxel (7.5-15 mg/kg，瘤内注射 IT，每周两次，持续六周；或每周 20-40 mg/kg，静脉注射 IV) 给药给携带 HNSCC 肿瘤 HN30 或 HN12 的 C57BL/6 小鼠。
结果：IT Docetaxel 提高了整体存活率和无病生存率，并逆转了肿瘤生长。在同等剂量水平下，IT Docetaxel 的肿瘤峰值浓度比 IV 治疗高 26 倍，肿瘤暴露时间比 IV 治疗长 24 倍。[4]

NCI-H460 cells (4 × 10^3) were grown in 100 μl of DMEM medium containing serum per well in a 96-well plate. After 24 h, the cells were treated with docetaxel (0, 0.2, 0.63, 2, 6.3, 20, 63 and 200 nmol/L, respectively) for 72 h. Every treatment was triplicate in the same experiment. Then 20 μl of MTS was added to each well for 1 to 4 h at 37°C. After incubation, the absorbance was read at a wavelength of 490 nm according to the manufacturer's protocol. The IC50 calculation was performed with GraphPad Prism 5.0 software [2].

Docetaxel (0, 10, 20, 30, 40, 60, and 80 mg/kg per week) was given once a week for 3 weeks for mice. Because more than 30 mg/kg per week of the drug caused body weight loss in mice, 20 mg/kg per week of docetaxel was judged to be the maximum nontoxic dose. Docetaxel (20 mg/kg per week) was given to mice once a week for 3 weeks at one of the following different points (2, 10, 14, or 22 HALO). Seventy-two hours after the final dosing of the agent, the intestinal mucosa of the small intestine (proximal 8 cm) was removed, fixed in 20 N Mildform solution (containing 8% formaldehyde in a buffered solution), and embedded in paraffin blocks, and sections of 5 mm were put on glass slides. Apoptosis was detected using the terminal deoxynucleotidyl transferase-mediated dUTP nick-end labeling (TUNEL) method, using the Apop Tag Peroxidase In Situ Apoptosis Detection Kit. Specimens were dewaxed and immersed in phosphate-buffered saline for 5 minutes at room temperature, incubated with 20 mg/ml proteinase K for 15 minutes at room temperature, and then quenched of endogenous peroxidase in 2% hydrogen peroxide in phosphate-buffered saline. Terminal deoxynucleotidyl transferase enzyme was applied directly onto the specimens, which were then incubated at 37°C for 1 hour. The reaction was terminated by transferring the slides to stop/wash buffer for 10 minutes at room temperature, and then specimens were covered with peroxidase-conjugated anti-digoxigenin antibody and incubated for 30 minutes at room temperature. Specimens were then soaked in staining buffer containing 0.05% diaminobenzidine to achieve color development. Finally, the specimens were counterstained by immersion in Mayer's hematoxylin solution. Apoptotic cells were counted under a light microscope in a good longitudinal crypt section. Starting at the base of the crypt column, the TUNEL-positive cells were counted up to the 18th cell position in each crypt.One hundred crypt sections were scored in each animal, and a frequency of TUNELpositive cells per crypt was calculated. Dosing time-dependent influence of docetaxel on intestinal apoptosis was also examined in female Balb/c mice [5].