2.3 – Cytotoxicity
ALK inhibitors (e.g., crizotinib) are used in clinical practice for
various ALK expressing malignancies therefore we hypothesized
that treatment with crizotinib would result in effective killing of
ALKfus cells13.
ALKfus+ Ba/F3 cells were plated in triplicate and
cultured with varying doses of Crizotinib (SelleckChem) in cytokine-free
media20. Cell viability and cell death were determined
with the Promega Cell Titer Glow assay after 48 hours in culture on a
BioTek Synergy H4 Hybrid plate reader via Gen5 2.1 Software; this was
repeated for ALKfus+ cells in the presence of IL-3.
Crizotinib-induced cytotoxicity in ALKfus+ Ba/F3 cells
was enhanced in the absence of IL-3 compared to cells cultured in the
presence of IL-3 (IC50 144nM vs. 1051nM for SPTBN1-ALK and 95nM
vs. 1277nM for RANBP2-ALK ) (Fig. 2C).