An investigation of reversal of imatinib resistance in the Bcr-Abl positive imatinib-resistant cell line K562r by dasatinib, nilotinib, rapamycin and bortezomib

Abstract

We investigated the second-generation tyrosine kinase inhibitors, dasatinib and nilotinib, for their potential to overcome resistance in the imatinib-resistant K562 cell line, and evaluated whether rapamycin, an mammalian target of rapamycin (mTOR) inhibitor, and bortezomib, a proteasome inhibitor, could increase imatinib sensitivity in resistant cell lines. Cell lines, including K562 (chronic myeloid leukaemia in blast crisis) and K562r (imatinib-resistant K562), were exposed to dasatinib, nilotinib, rapamycin, bortezomib, and rapamycin plus imatinib. Cell proliferation was measured by 3-[4, 5-dimethylthia-zol-2-yl]-2, 5-diphenyl tetrazolium bromide (MTT) assay and mTOR signaling pathways were assessed by Western blotting. Dasatinib, nilotinib, and bortezomib inhibited proliferation of K562 and K562r cell lines at nm concentrations. Resistance of K562r due to duplication of autophosphorylation of wild-type Bcr-Abl was not overcome by dasatinib and nilotinib, but was sensitive to bortezomib. Rapamycin partially inhibited proliferation of K562 and K562r cell lines. Rapamycin plus imatinib did not have a more inhibitory effect on the proliferation of K562 and K562r cell lines. K562r due to duplication of autophosphorylation of wild-type Bcr-Abl induced by imatinib was still partially resistant to dasatinib and nilotinib, but this was overcome by incremental dosing. Rapamycin did not enhance imatinib sensitivity. The blockade of the ubiquitin-proteasome pathway could be effective in overcoming resistance in the K562r imatinib-resistant cell line.