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.

PKCε:73 nM, PKCα:2 nM, PKA:15 nM, PhK:3 nM, CK1:163500 nM, S6K:5 nM, IR:61 nM, CK2:19500 nM, cdc2:9 nM, PKCγ:5 nM, EGFR:100 nM, CSK:2000 nM, ERK1:1500 nM, v-Src:6 nM, Ca 2+ /CaM PK-I1:20 nM, IGF-1R:6150 nM, PKCζ:1086 nM, PhK (Phosphorylase kinase):3 nM, TPK-IIB/Syk:16 nM, PKCη:4 nM, MLCK:21 nM, PKCδ:20 nM

METHODS: Human cervical cancer cells HeLa were treated with Staurosporine (1-10 nM) for 72 h, and cell viability was measured by MTT.
RESULTS: Staurosporine inhibited the proliferation of Hela cells in a dose-dependent manner, with an IC50 of about 10 nM. [1]
METHODS: Human pancreatic cancer cells PaTu 8988t and Panc-1 were treated with Staurosporine (1 μM) for 3-24 h, and cell death was detected by Flow Cytometry.
RESULTS: For PaTu 8988t cells, incubation with Staurosporine for 3-24 h significantly increased apoptosis and significantly decreased the number of viable cells; necrosis increased after 6-16 h. For Panc-1 cells, Staurosporine treatment significantly increased apoptosis and significantly decreased the number of viable cells after 9-24 h. The RESULTS were summarized as follows. [2]
METHODS: Human hepatocellular carcinoma cells HepG2 were treated with Staurosporine (20 nmol/L) for 6-24 h. The expression levels of target proteins were detected by Western Blot.
RESULTS: Staurosporine significantly inhibited the phosphorylation of mTOR and increased the expression of LC3-II, an autophagy marker protein, suggesting that Staurosporine activates autophagy effectively by inhibiting mTOR. [3]

METHODS: To assay anti-tumor activity in vivo, Staurosporine (3 mg/kg) and Lapatinib (50 mg/kg) were administered by gavage twice a week for two weeks to Nu/J-Foxn1 Nu/Nu mice harboring human mammary carcinoma tumors JIMT-1.
RESULTS: The combination of Staurosporine and Lapatinib inhibited tumor growth in a statistically significant manner. [4]
METHODS: To examine the effects on islet β-cell function, Staurosporine (0.4 mg/kg in 0.5% sodium carboxymethyl cellulose) was administered intraperitoneally to iPLA2β-/- C57BL6 mice once daily for two weeks. for two weeks.
RESULTS: Staurosporine impairs glucose tolerance and glucose-stimulated insulin secretion in pancreatic islets. [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)

H2O: < 0.1 mg/mL (insoluble)

DMSO: 31.56 mg/mL (67.65 mM), Sonication is recommended.

10% DMSO+40% PEG300+ 5% Tween 80 + 45% Saline: 3.16 mg/mL (6.77 mM), Solution.

10% DMSO+90% Saline: < 3.16 mg/mL (6.77 mM), Lower concentrations may be soluble, but exact solubility limit is unknown.

10% DMSO+40% PEG300+5% Tween 80+45% Saline: 3.1 mg/mL (6.64 mM), Solution.