Staurosporine (AM-2282) is a protein kinase inhibitor with ATP-competitive and non-selective inhibitory activity (IC50=6/15/2/3/3000 nM) against PKC, PKA, c-Fgr, phosphorylase kinase and TAOK2. Staurosporine also induces apoptosis.

PKCδ:20 nM, PKCγ:5 nM, PKCζ:1086 nM, PKCε:73 nM, PKCη:4 nM, PKCα:2 nM

方法：人宫颈癌细胞 HeLa 用 Staurosporine (1-10 nM) 处理 72 h，使用 MTT 方法检测细胞活力。
结果：Staurosporine 对 Hela 细胞增殖具有剂量依赖性抑制作用，IC50 约为 10 nM。[1]
方法：人胰腺癌细胞 PaTu 8988t、Panc-1 用 Staurosporine (1 μM) 处理 3-24 h，使用 Flow Cytometry 方法检测细胞死亡情况。
结果：PaTu 8988t 细胞，Staurosporine 孵育 3-24 h 显著增加了细胞凋亡率，并显著减少了生命细胞的数量；培养 6-16 h 后，坏死率增加。Panc-1 细胞，Staurosporine 处理在 9-24 h 后显著增加细胞凋亡并显著减少生命细胞的数量。[2]
方法：人肝癌细胞 HepG2 用 Staurosporine (20  nmol/L) 处理 6-24 h，使用 Western Blot 方法检测靶点蛋白表达水平。
结果：Staurosporine 显著抑制 mTOR 磷酸化并增加自噬标记蛋白 LC3-II 的表达，表明 Staurosporine 通过抑制 mTOR 有效激活自噬。[3]

方法：为检测体内抗肿瘤活性，将 Staurosporine (3 mg/kg) 和 Lapatinib (50 mg/kg) 灌胃给药给携带人乳腺癌肿瘤 JIMT-1 的 Nu/J-Foxn1 Nu/Nu 小鼠，每周两次，持续两周。
结果：Staurosporine 和 Lapatinib 的组合以统计学显著的方式抑制了肿瘤生长。[4]
方法：为检测对胰岛 β 细胞功能的影响，将 Staurosporine (0.4 mg/kg in 0.5% sodium carboxymethyl cellulose) 腹腔注射给 iPLA2β-/-的 C57BL6 小鼠，每天一次，持续两周。
结果：Staurosporine 会损害动物的糖耐量和胰岛的葡萄糖刺激的胰岛素分泌。[5]

Enzyme assay and binding assay: Protein kinase C is assayed in a reaction mixture (0.25 mL) containing 5 μmol of Tris/HCl, pH 7.5, 2.5 μmol of magnesium acetate, 50 μg of histone II S, 20 μg of phosphatidylserine, 0.88 μg of diolein, 125 nmol of CaCl2, 1.25 nmol of [γ-32]ATP (5-10 × 104 cpm/nmol) and 5 μg of partially purified enzyme. The binding of [3H]PDBu to protein kinase C is determined: Reaction mixture (200 μL contained 4 μmo1 of Tris/malate, pH 6.8, 20 μmol of KCl, 30 nmol of CaC12, 20 μg of phosphatidylserine, 5 μg of partially purified protein kinase C, 0.5% (final concentration) of DMSO,10 pmol of [3H]PDBu (l-3 × 104 cpm/pmol) and 10 μL of various amounts of Staurosporine.

Cells are exposed to Staurosporine for ~32 hours. Cells are fixed in 4% paraformaldehyde and stained with the DNA-binding dye Hoechst 33342. Cells are visualized under epifluorescence illumination, and the percentage of apoptotic cells (cells with condensed and fragmented DNA) is determined. (Only for Reference)