Erlotinib (Synonyms: NSC 718781)

Cell lines

BxPC-3 and MIAPaCa cells

Preparation Method

After the cells were incubated with Erlotinib for 72 h, the cell viability was detected by MTT assay.

Reaction Conditions

2 μM, 72 h

Applications

Erlotinib significantly inhibited BxPC-3 cell viability, but MIAPaCa cells were insensitive to Erlotinib.

Animal models

Orthotopic implantation model of BxPC-3 cells in mice

Preparation Method

Mice were randomized into the following treatment groups (n = 7): (a) untreated control; (b) only B-DIM (50 mg/kg body weight), intragastric once every day; (c) Erlotinib (50 mg/kg body weight), everyday i.p. for 15 days; and (d) B-DIM and Erlotinib. All mice were killed on day 3 following last dose of treatment, and their body weight was determined.

Dosage form

50 mg/kg/day, 15 days, i.p.

Applications

Compared with tumors in the control group, the tumor weight of mice in the Erlotinib group was reduced by 35%.

Preparation Method

The kinase reaction was performed in50 μl of 50 mM HEPES (pH 7.3), containing 125 mM NaCl, 24 mM MgCl₂, 0.1 mM sodium orthovanadate, 20 μM ATP, 1.6 μg/ml EGF, and 15 ng of EGFR. The Erlotinib in DMSO was added to give a final DMSO concentration of 2.5%. Phosphorylation was initiated by addition of ATP and proceeded for 8 min at room temperature with constant shaking. The kinase reaction was terminated by aspiration of the reaction mixture and was washed 4 times with wash buffer. Phosphorylated POT was measured by 25 min of incubation with 50 μl per well HRP-conjugated PY54 antiphosphotyrosine antibody, diluted to 0.2 μg/mI in blocking buffer (3% BSA and 0.05% Tween 20 in PBS). Removed antibody by aspiration and washed the plate 4 times with wash buffer. The colonmetric signal was developed by addition of TMB Microwell Peroxidase Substrate, 50 μl per well, and stopped by the addition of 0.09 M sulfuric acid, 50 μ1 per well. Phosphotyrosine is estimated by measurement of absorbance at 450 nm.

Reaction Conditions

0.01-1000 nM

Applications

Erlotinib is a directly acting inhibitor of human EGFR tyrosine kinase with an IC₅₀ of 2 nM and reduces EGFR autophosphorylation in intact tumor cells with an IC₅₀ of 20 nM. 

References:
[1] Ali S, Banerjee S, Ahmad A, et al. Apoptosis-inducing effect of erlotinib is potentiated by 3, 3′-diindolylmethane in vitro and in vivo using an orthotopic model of pancreatic cancer[J]. Molecular cancer therapeutics, 2008, 7(6): 1708-1719.
[2] Moyer J D, Barbacci E G, Iwata K K, et al. Induction of apoptosis and cell cycle arrest by CP-358,774, an inhibitor of epidermal growth factor receptor tyrosine kinase[J]. Cancer research, 1997, 57(21): 4838-4848.