Abstract
In this study, we examined the role of JAK/STAT signaling in the regulation of chronic leukemia K562 cell proliferation. STAT3 and STAT5 tyrosine phosphorylation was used as a marker of the activation status of STAT proteins. We demonstrated that, in growing cultures of K562 cells, both STAT3 and STAT5 are constitutively activated. To determine the significance of STAT activity in maintaining the high level of K562 proliferation, we tested two JAK inhibitors, AG-490 (JAK2 and JAK3 inhibitor) and WHI-P131 (a new specific JAK3 inhibitor). We showed that, during the prolonged cultivation (48 h) of K562 cells with AG-490 or WHI-P131, the cells remained viable. It was found that treatment with WHI-P131 (30–100 μM) decreased tyrosine phosphorylation of STAT5 and did not affect the high level of STAT3 phosphorylation. In proliferating K562 cells, AG-490 (25–50 μM) did not influence on STAT3 and STAT5 phosphorylation. The flow cytometry analysis revealed a dose-dependent decrease in G1 and S phases and an increase in G2/M phases in WHI-P131-treated K562 cells and no changes in cell cycle structure in AG-490-treated cultures. Thus, our findings indicate the preferential role of STAT5 (not constitutively active STAT3) in the proliferation of leukemia K562 cells and demonstrate the specificity of WHI-P131 inhibitory effect; unlike other JAK drugs that stimulate apoptosis and decrease proliferation, WHI-P131 prevents K562 cells growth by arresting in G2/M phases of the cell cycle.
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Abbreviations
- IL-2:
-
interleukin-2
- PBL:
-
peripheral blood lymphocytes
- JAK:
-
Janus kinase
- STAT:
-
signal transduser and activator of transcription
References
Alexander, W.S. and Hilton, D.J., The Role of Suppressors of Cytokine Signaling (SOCS) Proteins in Regulation of the Immune Response. Annu. Rev. Immunol., 2004, vol. 22, pp. 503–529.
Alvarez, J.V. and Frank, D.A., Genome-Wide Analysis of STAT Target Genes: Elucidating the Mechanism of STAT-Mediated Oncogenesis. Cancer Biol. Ther., 2004, vol. 3, pp. 1045–1050.
Amin, H.M., Medeiros, L.J., Ma, Y., Feretzaki, M., Das, P., Leventaki, V., Rassidakiss, G.Z., O’Connor, S.L., McDonnell, T.J., and Lai, R., Inhibition of JAK3 Induces Apoptosis and Decreases Anaplastic Lymphoma Kinase Activity in Anaplastic Large Cell Lymphoma. Oncogene, 2003, vol. 22, pp. 5399–5407.
Amin, H.V., McDonnell, T.J., Ma, Y., Lin, Q., Fujio, Y., Kunisada, K., Leventaki, V., Das, P., Rassidakis, G.Z., Cutler, C., Medeiros, L.J., and Lai, R., Selective Inhibition of STAT3 Induces Apoptosis and G(1) Cell Cycle Arrest in ALK-Positive Anaplastic Large Cell Lymphoma. Oncogene, 2004, vol. 23, pp. 5399–407.
Baus, D. and Pfitzner, E., Specific Function of STAT3, SOCS1, and SOCS3 Is the Regulation of Proliferation and Survival of Classical Hodgkin Lymphoma Cells. Int. J. Cancer, 2006, vol. 118, pp. 1404–1413.
Berridge, M.J., Lymphocyte Activation in Health and Disease. Critical Rev. Immunol., 1997, vol. 17, pp. 155–178.
Bradford, M.M., A Rapid and Sensitive Method for the Quantitation of Microgram Quantities of Protein Utilizing the Principle of Protein-dye Binding. Anal. Biochem., 1976, vol. 72, pp. 248–254.
Cochet, O., Frelin, C., Peyron, J.F., and Imbert, V., Constitutive Activation of STAT Proteins in the HDLM-2 and L540 Hodgkin Lymphoma-Derived Cell Lines Supports Cell Survival. Cell Signal., 2006, vol. 18, pp. 449–455.
Darnell, J.E., Jr., STATs and Gene Regulation. Science, 1997, vol. 277, pp. 1630–1635.
Ferrojoli, A., Faderi, S., Van, Q., Koch, P., Harris, D., Liu, Z., Hazan-Halevy, I., Wang, Y., Katarjian, H.M., Priebe, W., and Wstron, Z., WP12066 Disrupts Janus Kinase-2 and Induces Caspase-Dependent Apoptosis in Acute Muelogenous Leukemia Cells. Cancer. Res., 2007, vol. 67, pp. 11291–11299.
Khwaja, A., The Role of Janus Kinases in Haemopoiesis and Haematological Malignancy. Br. J. Haematol., 2006, vol. 134, pp. 366–384.
Kim, B.H., Yin, C.H., Cuo, Q., Bach, E.A., Lee, H., Sandoval, C., Jayabose, S., Ulaczyk-Lesanko, A., Hall, D.G., and Baeg, G.H., A Small-Molecule Compound Identified through a Cell-Based Screening Inhibits JAK/STAT Pathway Signaling in Human Cancer Cells. Mol. Cancer Ther., 2008, vol. 7, pp. 2672–2780.
Kirken, R.A., Erwin, R.A., Wang, L., Wang, Y., Rui, H., and Farrar, W.L., Functional Uncoupling of the Janus-Kinase 3-STAT5 Pathway in Malignant Growth of Human T Cell Leukemia Virus Type 1-Transformed Human T-Cells. Immunology, 2000, vol. 165, pp. 5097–5104.
Kube, D., Holtick, U., Vockerodt, M., Ahmadi, T., Haier, B., Behrmann, I., Heinrich, P.C., Diehl, V., and Tesch, H., STAT3 Is Constitutively Activated in Hodgkin Cell Lines. Blood, 2001, vol. 98, pp. 762–770.
Leonard, W.J., Cytokines and Immunodeficiency Diseases. Immunology, 2001, vol. 1, pp. 200–208.
Leonard, W.J. and Lin, J.-X., Cytokine Receptor Signaling Pathways. J. Allergy Clin. Immunol., 2000, vol. 105, pp. 877–888.
Lewis, R.S. and Ward, A.C., STAT5 as a Diagnostic Marker for Leukemia. Exp. Rev. Mol. Diagn., 2008, vol. 8, pp. 73–82.
Lin, J.-X. and Leonard, W.J., The Role of STAT5a and STAT5b in Signaling by IL-2 Family Cytokines. Oncogene, 2000, vol. 19, pp. 2566–2576.
Nagy, Z.S., Rui, H., Stepkowski, S.M., Karras, J., and Kirken, R.A., A Preferential Role of STAT5, Not Costitutively Active STAT3, in Promoting Survival of a Human Lymphoid Tumor. J. Immunol., 2006, vol. 177, pp. 5032–5040.
Ozawa, Y., Williams, A.H., Estes, M.L., Matsushita, N., Boschelli, F., Jove, R., and List, A.F., Src Family Kinases Promote AML Cell Survival through Activation of Signal Transducers and Activators of Transcription (STAT). Leuk Res., 2008, vol. 32, pp. 893–903.
Pesu, M., Candotti, F., Husa, M., Hofmann, S.R., Notarangelo, L.D., and O’shea, J.J., JAK3, Severe Combined Immunodeficiency, and a New Class of Immunosuppressive Drugs. Immunol. Rev., 2005, vol. 203, pp. 127–142.
Rane, S.G. and Reddy, E.P., Janus Kinases: Components of Multiple Signaling Pathways. Oncogene, 2000, vol. 19, pp. 5662–5679.
Sudbeck, E.A., Liu, X.P., Narla, R.K., Mahajan, S., Ghosh, S., Mao, C., and Uckin, F.M., Structure-Based Design of Specific Inhibitors of Janus Kinase 3 as Apoptosis-Inducing Antileukemic Agents. Clin. Cancer Res., 1999, vol. 5, pp. 1569–1582.
Tong, H., Ren, Y., Zhang, F., and Jin, J., Homoharringtonine Affects the JAK2-STAT5 Signal Pathway through Alteration of Protein Tyrosine Kinase Phosphorylation in Acute Myeloid Leukemia Cells. Eur. J. Haematol., 2008, vol. 81, pp. 259–266.
Turkson, J. and Jone, R., STAT Proteins: Novel Molecular Targets for Cancer Drug Discovery. Oncogene, 2000, vol. 19, pp. 6613–6626.
Vereninov, A.A., Vinogradova, T.A., Ivakhnyuk, I.S., Marakhova, I.I., and Toropova, F.V., The Use of Flame-Emission Analysis for Measuring Alkaline Cation Flux across Cell Membrane. Tsitologiya, 1982, vol. 24, no. 1, pp. 98–103.
Wang, L.H., Kirken, R.A., Erwin, R.A., Yu, C-R., and Farrar, W.L., JAK3, STAT, and MAPK Signalling Pathways as Novel Molecular Targets for the Tyrphostin AG-490 Regulation of IL-2-Mediated T Cell Response. J. Immunol., 1999, vol. 162, pp. 3897–3904.
Yamaoka K., Saharinen, P., Pesu, M., Holt, V.E., 3rd, Silvennoinen, O., and O’shea, J.J., The Janus Kinases (Jaks). Genome Biol., 2004, vol. 5, pp. 253–259.
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Original Russian Text © E.V. Mityushova, N.D. Aksenov, I.I. Marakhova, 2010, published in Tsitologiya, Vol. 52, No. 2, 2010, pp. 184–189.
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Mityushova, E.V., Aksenov, N.D. & Marakhova, I.I. STAT5 in regulation of chronic leukemia K562 cell proliferation: Inhibitory effect of WHI-P131. Cell Tiss. Biol. 4, 63–69 (2010). https://doi.org/10.1134/S1990519X10010062
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DOI: https://doi.org/10.1134/S1990519X10010062