ISSN 1214-0287 (on-line)
ISSN 1214-021X (printed)

Volume 4 (2006), No 2, p 95-104

Imatinib mesylate affects tyrosine kinase activity in both leukemic and normal primary mononuclear blood cells

Katerina Kuzelova, Dana Grebenova, Michaela Pluskalova, Iuri Marinov, Hana Klamova, Zbynek Hrkal

Address: Katerina Kuzelova, Institute of Hematology and Blood Transfusion, U Nemocnice 1, 128 20 Praha 2, Czech Republic

Received 9th December 2005.
Revised 14th January 2006.
Published online 4th April 2006.

Full text article (pdf)

Tyrosine kinase (TK) activity in primary mononuclear blood cells (MNBC) derived from chronic myelogenous leukemia (CML) patients in the chronic phase as well as from healthy donors was measured by a sensitive time-resolved fluorescence method using the Delfia Tyrosine Kinase kit. The level of phosphotyrosine was assessed in parallel by flow-cytometry. The experimental protocol for Delfia was optimized using a K562 cell line. A large part (20 to 50%) of the fluorescence signal from K562 cells was sensitive to Imatinib mesylate, an inhibitor of Bcr-Abl tyrosine kinase, which is currently the leading drug in CML treatment. In primary MNBC, the direct contribution of Bcr-Abl itself to the signal was low. However, a 48h treatment of MNBC with 5 microM Imatinib resulted in a significant reduction of the observed TK activity (mean TK activity value: 56% of control) paralleled by a decrease in the phosphotyrosine level in the CML group. Modification of TK activity by Imatinib was observed also in the donor group. Imatinib mesylate thus probably affects cell signalization even in Bcr-Abl negative cells.

tyrosin-kinase activity; Imatinib; K562; primary blood cells; leukemia

Appel S, Boehmler AM, Grunebach F et al.: Imatinib mesylate affects the development and function of dendritic cells generated from CD34+ peripheral blood progenitor cells. Blood 103:538-544, 2004.

Bartolovic K, Balabanov S, Hartmann U et al.: Inhibitory effect of imatinib on normal progenitor cells in vitro. Blood 103:523-529, 2004.

Baskaynak G, Kreuzer KA, Schwarz M et al.: Squamous cutaneous epithelial cell carcinoma in two CML patients with progressive disease under imatinib treatment. Eur J Haematol 70:231-234, 2003.

Calabretta B, Perrotti D: The biology of CML blast crisis. Blood 103:4010-4022, 2004.

Cwynarski K, Laylor R, Macchiarulo E et al.: Imatinib inhibits the activation and proliferation of normal T lymphocytes in vitro. Leukemia 18:1332-1339, 2004.

Deininger MWN, Goldman JM, Lydon N, Melo JV: The tyrosine kinase inhibitor CGP57148B selectively inhibits the growth of BCR-ABL-positive cells. Blood 90:3691-3698, 1997.

Deininger M, Buchdunger E, Druker BJ: The development of imatinib as a therapeutic agent for chronic myeloid leukemia. Blood 105:2640-2653, 2005.

Dewar AL, Cambareri AC, Zannettino ACW et al.: Macrophage colony-stimulating factor receptor c-fms is a novel target of imatinib. Blood 105:3127-3132, 2005.

Dewar AL, Domaschenz RM, Doberty KV et al.: Imatinib inhibits the in vitro development of the monocyte/macrophage lineage from normal human bone marrow progenitors. Leukemia 17:1713-1721, 2003.

Dong R, Cwynarski K, Entwistle A et al.: Dendritic cells from CML patients have altered actin organization, reduced antigen processing and impaired migration. Blood 101:3560-3567, 2003.

Druker BJ, Tamura S, Buchdunger E et al.: Effects of a selective inhibitor of the Abl tyrosine kinase on the growth of Bcr-Abl positive cells. Nat Med 2:561-566, 1996.

Fang G, Kim CN, Perkins CL et al.: CGP57148B (STI-571) induces differentiation and apoptosis and sensitizes Bcr-Abl-positive human leukemia cells to apoptosis due to antileukemic drugs. Blood 96:2246-2253, 2000.

Ferrao PT, Frost MJ, Siah SP, Ashman LK: Overexpression of P-glycoprotein in K562 cells does not confer resistance to the growth inhibitory effects of imatinib (STI571) in vitro. Blood 102:4499-4503, 2003.

Gambacorti-Passerini C, Le Coutre P, Mologni L et al.: Inhibition of the ABL kinase activity blocks the proliferation of BCR/ABL+ leukemic cells and induces apoptosis. Blood Cells Mol Dis 23:380-394, 1997.

Grebenova D, Kuzelova K, Smetana K et al.: Mitochondrial and endoplasmic reticulum stress-induced apoptotic pathways are activated by 5-aminolevulinic acid-based photodynamic therapy in HL60 leukemia cells. J Photochem Photobiol B, Biol 69:71-85, 2003.

Guo JQ, Lian JY, Xian YM et al.: BCR-ABL Protein expression in peripheral blood cells of chronic myelogenous leukemia patients undergoing therapy. Blood 83:3629-3637, 1994.

Holtz MS, Slovak ML, Zhang F et al.: Imatinib mesylate (STI571) inhibits growth of primitive malignant progenitors in chronic myelogenous leukemia through reversal of abnormally increased proliferation. Blood 99:3792-3800, 2002.

Jacquel A, Herrant M, Legros L et al.: Imatinib induces mitochondria-dependent apoptosis of the Bcr-Abl positive K562 cell line and its differentiation towards the erythroid lineage. FASEB J 17:2160-2162, 2003.

Kantarjian H, Talpaz M, O´Brien S et al.: Highdose imatinib therapy in newly diagnosed Philadelphia chromosome-positive chronic phase chronic myeloid leukemia. Blood 103:2873-2878, 2004.

Kawano T, Horiguchi-Yamada J, Saito S et al.: Ectopic cyclin D1 expression blocks STI571-induced erythroid differentiation of K562 cells. Leuk Res 28:623-629, 2004.

Kotaki M, Motoji T, Takanashi M et al.: Antiproliferative effect of the abl tyrosine kinase inhibitor STI571 on the P-glycoprotein positive K562/ADM cell line. Cancer Lett 199:61-68, 2003.

Krystal GW: Imatinib mesylate (STI571) for myeloid malignancies other than CML. Leuk Res 28S1:S53-S59, 2004.

Kuzelova K, Grebenova D, Marinov I, Hrkal Z: Fast apoptosis and erythroid differentiation induced by Imatinib mesylate in JURL-MK1 cells. J Cell Biochem 95:268-280, 2005.

Le Coutre P, Kreuzer KA, Pursche S et al.: Pharmacokinetics and cellular uptake of imatinib and its main metabolite CGP74588. Cancer Chemother Pharmacol 53:313-323, 2004.

Mahon FX, Deininger MWN, Schultheis B et al.: Selection and characterization of BCR-ABL positive cell lines with differential sensitivity to the tyrosine kinase inhibitor STI571: diverse mechanisms of resistance. Blood 96:1070-1079, 2000.

Mahon FX, Belloc F, Lagarde V et al.: MDR1 gene overexpression confers resistance to imatinib mesylate in leukemia cell line models. Blood 101:2368-2373, 2003.

Manley PW, Cowan-Jacob SW, Buchdunger E et al.: Imatinib: a selective tyrosine kinase inhibitor. Eur J Cancer 38:S19-S27, 2002.

Mattiuzzi GN, Cortes JE, Talpaz M et al.: Development of Varicella-Zoster virus infection in patients with chronic myelogenous leukemia treated with imatinib mesylate. Clin Cancer Res 9:976-980, 2003.

Mohty M, Jourdan E, Mami NB et al.: Imatinib and plasmacytoid dendritic cell function in patients with chronic myeloid leukemia. Blood 103:4666-4668, 2004.

Mukai M, Che XF, Furukawa T et al.: Reversal of the resistance to STI571 in human chronic myelogenous leukemia K562 cells. Cancer Sci 94:557-563, 2003.

Rodley P, McDonald M, Price B et al.: Comparative genomic hybridization reveals previously undescribed amplifications and deletions in the chronic myeloid leukemiaderived K-562 cell line. Genes Chromosomes Cancer 19:36-42, 1997.

Sawyers C.L., Hochhaus A., Feldman E. et al.: Imatinib induces hematological and cytogenetic responses in patients with chronic myelogenous leukemia in myeloid blast crisis: results of a phase II study. Blood 99:3530-3539, 2002.

Shaul Y, Ben-Yehoyada M: Role of c-Abl in the DNA damage stress response. Cell Res 15:33-35, 2005.

Steelman LS, Pohnert SC, Shelton JG et al.: JAK/STAT, RaF/MEK/ERK, PI3K/Akt and BCR-ABL in cell cycle progression and leukemogenesis. Leukemia 18:189-218, 2004.

Taieb J, Maruyama K, Borg C et al.: Imatinib mesylate impairs Flt3L-mediated dendritic cell expansion and antitumor effects in vivo. Blood 103:1966-1967, 2004.

Takahashi N, Miura I, Saitoh K, Miura AB: Lineage involvement of stem cells bearing the Philadelphia chromosome in chronic myeloid leukemia in the chronic phase as shown by a combination of fluorescence-activated cell sorting and fluorescence in situ hybridization. Blood 92:4758-4763, 1998.

Talpaz M, Silver RT, Druker BJ et al.: Imatinib induces durable hematologic and cytogenetic responses in patients with accelerated phase chronic myeloid leukemia: results of a phase 2 study. Blood 99:1928-1937, 2002.

Traina F, Carvalheira JBC, Saad MJA et al.: BCR-ABL binds to IRS-1 and IRS-1 phosphorylation is inhibited by imatinib in K562 cells. FEBS Lett 535:17-22, 2003.

Weisberg E, Griffin JD: Mechanism of resistance to the ABL tyrosine kinase inhibitor STI571 in BCR/ABL-transformed hematopoietic cell lines. Blood 95:3498-3505, 2000.

Wu SQ, Voelkerding KV, Sabatini L et al.: Extensive amplification of bcr/abl fusion genes clustered on three marker chromosomes in human leukemic cell line K-562. Leukemia 9:858-862, 1995.

Kuzelova K, Pluskalova M, Brodska B, Otevrelova P, Elknerova K, Grebenova D, Hrkal Z: Suberoylanilide Hydroxamic Acid (SAHA) at Subtoxic Concentrations Increases the Adhesivity of Human Leukemic Cells to Fibronectin. J Cell Biochem 109:184-195, 2010.