Imatinib mesylate affects extracellular ATP catabolism and expression of NTPDases in a chronic myeloid leukemia cell line

Abstract

Chronic myeloid leukemia (CML) is a myeloproliferative neoplasm, characterized by the occurrence of the t(9;22)(q34;q11) translocation. First-line therapy for CML consists of treatment with imatinib mesylate, which selectively inhibits the BCR-ABL protein by competing for its ATP-binding site. Adenine nucleotide signaling is modulated by the ectonucleotidases and this pathway is related to tumorigenic processes. Considering the relationship between ATP and cancer, we aimed to evaluate the influence of imatinib mesylate on the expressions and functions of the NTPDase and ecto-5′-nucleotidase (CD73) enzymes in imatinib-sensitive and -resistant K-562 cell lines. mRNA analysis showed that K-562 cells express all ENTPDs and NT5E. However, when treated with imatinib mesylate for 24 h, the expression of ENTPD1, -2, -3 and -5 increased, leading to a higher nucleotides hydrolysis rate. HPLC analysis identified increased ATP degradation in cells after 24 h of treatment, with consequent ADP and AMP formation, corroborating the increase in gene and protein expression of ectonucleotidases as observed in previous results. On the other hand, we observed that imatinib-resistant K-562 cells presented a decrease in nucleotide hydrolysis and expressions of ENTPD1 and -5. These results suggest an involvement of imatinib in modulating ectonucleotidases in CML that will need further investigation. Since these ectonucleotidases have important catalytic activities in the tumor microenvironment, their modulation in CML cells may represent an important therapeutic approach to regulate levels of extracellular adenine nucleotides.

Betulinic Acid-Brosimine B Hybrid Compound Has a Synergistic Effect with Imatinib in Chronic Myeloid Leukemia Cell Line, Modulating Apoptosis and Autophagy

“…However, the appearance of adverse effects elevated cases of treatment discontinuation. The main adverse reactions involving imatinib are gastrointestinal disturbances and rashes, in addition to thrombocytopenia and neutropenia [18][19][20]. Importantly, the discontinuation of treatment may lead to the development of therapeutic resistance; resistance to imatinib is a complex process without a defined cause.…”

“…Cells were cultured in RPMI-1640 medium with 10% fetal bovine serum and 100 U/mL pen-streptomycin and incubated at 37 • C in 5% CO 2 . The K-562R (imatinib-resistant cell line) was generated as described by Willig et al [18]. Peripheral blood mononuclear cells (PBMNCs) were used as control and were obtained by centrifuging blood from healthy donors over a Histopaque®-1077 (Sigma-Aldrich, St. Louis, MO, USA) density gradient.…”

Identification of Novel Genes and Associated Drugs in Cervical Cancer by Bioinformatics Methods

“…Regorafenib is a novel multikinase inhibitor that targets tumor genesis, tumor angiogenesis, and maintenance of signaling in the tumor microenvironment by inhibiting multiple protein kinases that promote tumor growth [ 43 ]. Dasatinib [ 44 ], Ponatinib [ 45 ], and ImatinibJournal [ 46 ] are tyrosine kinase inhibitors that effectively lock tyrosine kinase into inactive conformation by binding to highly conserved ATP binding sites, thereby inhibiting BCR-ABL activity. They can inhibit multiple oncogenic pathways activated by BCR-ABL tyrosine kinase and promote cell proliferation and survival, including the MAPK/ERK pathway, JAK-Stat pathway, and PI3K/Akt pathway.…”

Augmented Therapeutic Potential of EC-Synthetic Retinoids in Caco-2 Cancer Cells Using an In Vitro Approach

“…All cellular ATPases were extracted from Caco-2 cells after lysis with 1% Triton X-100, 5 mM EDTA, 50 mM NaCl and 20 mM HEPES (pH 7.4). To normalize the total protein content in the sample, the concentration of all proteins in the sample was measured by a Bradford assay [ 150 ], and 200-µg total proteins were used per each ml of cell lysate [ 151 ]. For the assessment of the calcium-independent ATPase activity, the reactions were performed in the presence of 200 µM EDTA in addition to 1 mM ATP, 20 mM sodium azide and 50 mM Tris (pH 7.5) [ 148 ].…”