Asian Pacific Journal of Cancer Prevention

Asian Pacific Journal of Cancer Prevention (아시아태평양암예방학회)
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Silencing of Suppressor of Cytokine Signaling-3 due to Methylation Results in Phosphorylation of STAT3 in Imatinib Resistant BCR-ABL Positive Chronic Myeloid Leukemia Cells

  • Al-Jamal, Hamid AN (Department of Haematology, School of Medical Sciences, Universiti Sains Malaysia) ;
  • Jusoh, Siti Asmaa Mat (Department of Haematology, School of Medical Sciences, Universiti Sains Malaysia) ;
  • Yong, Ang Cheng (Department of Haematology, School of Medical Sciences, Universiti Sains Malaysia) ;
  • Asan, Jamaruddin Mat (Department of Immunology, School of Medical Sciences, Universiti Sains Malaysia) ;
  • Hassan, Rosline (Department of Haematology, School of Medical Sciences, Universiti Sains Malaysia) ;
  • Johan, Muhammad Farid (Department of Haematology, School of Medical Sciences, Universiti Sains Malaysia)
  • Published : 2014.06.15
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Abstract

Background: Silencing due to methylation of suppressor of cytokine signaling-3 (SOCS-3), a negative regulator gene for the JAK/STAT signaling pathway has been reported to play important roles in leukemogenesis. Imatinib mesylate is a tyrosine kinase inhibitor that specifically targets the BCR-ABL protein and induces hematological remission in patients with chronic myeloid leukemia (CML). Unfortunately, the majority of CML patients treated with imatinib develop resistance under prolonged therapy. We here investigated the methylation profile of SOCS-3 gene and its downstream effects in a BCR-ABL positive CML cells resistant to imatinib. Materials and Methods: BCR-ABL positive CML cells resistant to imatinib (K562-R) were developed by overexposure of K562 cell lines to the drug. Cytotoxicity was determined by MTS assays and $IC_{50}$ values calculated. Apoptosis assays were performed using annexin V-FITC binding assays and analyzed by flow cytometry. Methylation profiles were investigated using methylation specific PCR and sequencing analysis of SOCS-1 and SOCS-3 genes. Gene expression was assessed by quantitative real-time PCR, and protein expression and phosphorylation of STAT1, 2 and 3 were examined by Western blotting. Results: The $IC_{50}$ for imatinib on K562 was 362nM compared to 3,952nM for K562-R (p=0.001). Percentage of apoptotic cells in K562 increased upto 50% by increasing the concentration of imatinib, in contrast to only 20% in K562-R (p<0.001). A change from non-methylation of the SOCS-3 gene in K562 to complete methylation in K562-R was observed. Gene expression revealed down-regulation of both SOCS-1 and SOCS-3 genes in resistant cells. STAT3 was phosphorylated in K562-R but not K562. Conclusions: Development of cells resistant to imatinib is feasible by overexposure of the drug to the cells. Activation of STAT3 protein leads to uncontrolled cell proliferation in imatinib resistant BCR-ABL due to DNA methylation of the SOCS-3 gene. Thus SOCS-3 provides a suitable candidate for mechanisms underlying the development of imatinib resistant in CML patients.

Keywords

References

  1. A J, Qian S, Wang G, et al (2010). Chronic myeloid leukemia patients sensitive and resistant to imatinib treatment show different metabolic responses. PLoS One, 5, 13186. https://doi.org/10.1371/journal.pone.0013186
  2. Bar-Natan M, Nelson EA, Xiang M, Frank DA (2012). STAT signaling in the pathogenesis and treatment of myeloid malignancies. JAKSTAT, 1, 55-64.
  3. Benekli M, Baumann H, Wetzler M (2009). Targeting signal transducer and activator of transcription signaling pathway in leukemias. J Clin Oncol, 27, 4422-32. https://doi.org/10.1200/JCO.2008.21.3264
  4. Benekli M, Xia Z, Donohue KA, et al (2002). Constitutive activity of signal transducer and activator of transcription 3 protein in acute myeloid leukemia blasts is associated with short disease-free survival. Blood, 99, 252-7. https://doi.org/10.1182/blood.V99.1.252
  5. Binato R, Mencalha A, Pizzatti L, et al (2009). RUNX1T1 is overexpressed in imatinib mesylate-resistant cells. Mol Med Rep, 2, 657-61.
  6. Branford S, Rudzki Z, Harper A, et al (2003). Imatinib produces significantly superior molecular responses compared to interferon alfa plus cytarabine in patients with newly diagnosed chronic myeloid leukemia in chronic phase. Leukemia, 17, 2401-9. https://doi.org/10.1038/sj.leu.2403158
  7. Capello D, Deambrogi C, Rossi D, et al (2008). Epigenetic inactivation of suppressors of cytokine signalling in Philadelphia-negative chronic myeloproliferative disorders. Br J Haematol, 141, 504-11. https://doi.org/10.1111/j.1365-2141.2008.07072.x
  8. Catlett-Falcone R, Landowski TH, Oshiro MM, et al (1999). Constitutive activation of Stat3 signaling confers resistance to apoptosis in human U266 myeloma cells. Immunity, 10, 105-15. https://doi.org/10.1016/S1074-7613(00)80011-4
  9. Coley HM (2004). Development of drug-resistant models. Methods Mol Med, 88, 267-73.
  10. Croker BA, Kiu H, Nicholson SE (2008). SOCS regulation of the JAK/STAT signalling pathway. Semin Cell Dev Biol, 19, 414-22. https://doi.org/10.1016/j.semcdb.2008.07.010
  11. Druker BJ, Guilhot F, O'Brien SG, et al (2006). Five-year follow-up of patients receiving imatinib for chronic myeloid leukemia. N Engl J Med, 355, 2408-17. https://doi.org/10.1056/NEJMoa062867
  12. Esposito N, Colavita I, Quintarelli C, et al (2011). SHP-1 expression accounts for resistance to imatinib treatment in Philadelphia chromosome-positive cells derived from patients with chronic myeloid leukemia. Blood. 118, 3634-44. https://doi.org/10.1182/blood-2011-03-341073
  13. Frohling S, Scholl C, Levine RL, et al (2007). Identification of driver and passenger mutations of FLT3 by high-throughput DNA sequence analysis and functional assessment of candidate alleles. Cancer cell, 12, 501-13. https://doi.org/10.1016/j.ccr.2007.11.005
  14. Furqan M, Mukhi N, Lee B, Liu D (2013). Dysregulation of JAK-STAT pathway in hematological malignancies and JAK inhibitors for clinical application. Biomark Res, 1, 5. https://doi.org/10.1186/2050-7771-1-5
  15. Gruber FX, Ernst T, Porkka K, et al (2012). Dynamics of the emergence of dasatinib and nilotinib resistance in imatinib-resistant CML patients. Leukemia, 26, 172-7. https://doi.org/10.1038/leu.2011.187
  16. Herman JG, Graff JR, Myohanen S, Nelkin BD, Baylin SB (1996). Methylation-specific PCR: a novel PCR assay for methylation status of CpG islands. Proc Natl Acad Sci USA, 93, 9821-6. https://doi.org/10.1073/pnas.93.18.9821
  17. Ihle JN (1995). The Janus protein tyrosine kinase family and its role in cytokine signaling. Adv Immunol, 60, 1-35. https://doi.org/10.1016/S0065-2776(08)60582-9
  18. Jatiani SS, Baker SJ, Silverman LR, Reddy EP (2010). Jak/STAT pathways in cytokine signaling and myeloproliferative disorders: approaches for targeted therapies. Genes Cancer, 1, 979-93. https://doi.org/10.1177/1947601910397187
  19. Kubo M, Hanada T, Yoshimura A (2003). Suppressors of cytokine signaling and immunity. Nat Immunol, 4, 1169-76. https://doi.org/10.1038/ni1012
  20. Liang P, Cheng SH, Cheng CK, et al (2013). Platelet factor 4 induces cell apoptosis by inhibition of STAT3 via up-regulation of SOCS3 expression in multiple myeloma. Haematologica, 98, 288-95. https://doi.org/10.3324/haematol.2012.065607
  21. Liu P, Wang X, Hu C, Hu T (2011). Inhibition of proliferation and induction of apoptosis by trimethoxyl stilbene (TMS) in a lung cancer cell line. Asian Pac J Cancer Prev, 12, 2263-9.
  22. Liu TC, Lin SF, Chang JG, et al (2003). Epigenetic alteration of the SOCS1 gene in chronic myeloid leukaemia. Br J Haematol, 123, 654-61. https://doi.org/10.1046/j.1365-2141.2003.04660.x
  23. Melo JV, Barnes DJ (2007). Chronic myeloid leukaemia as a model of disease evolution in human cancer. Nat Rev Cancer, 7, 441-53. https://doi.org/10.1038/nrc2147
  24. Morgillo F, Martinelli E, Troiani T, et al (2011). Antitumor activity of sorafenib in human cancer cell lines with acquired resistance to EGFR and VEGFR tyrosine kinase inhibitors. PLoS One, 6, 28841. https://doi.org/10.1371/journal.pone.0028841
  25. Niwa Y, Kanda H, Shikauchi Y, et al (2005). Methylation silencing of SOCS-3 promotes cell growth and migration by enhancing JAK/STAT and FAK signalings in human hepatocellular carcinoma. Oncogene, 24, 6406-17.
  26. Qin S, Ai F, Ji WF, et al (2013). miR-19a promotes cell growth and tumorigenesis through targeting SOCS1 in gastric cancer. Asian Pac J Cancer Prev, 14, 835-40. https://doi.org/10.7314/APJCP.2013.14.2.835
  27. Qiu X, Guo G, Chen K, et al (2012). A requirement for SOCS-1 and SOCS-3 phosphorylation in Bcr-Abl-induced tumorigenesis. Neoplasia, 14, 547-58. https://doi.org/10.1596/neo.12230
  28. Radich JP, Dai H, Mao M, et al (2006). Gene expression changes associated with progression and response in chronic myeloid leukemia. Proc Natl Acad Sci USA, 103, 2794-9. https://doi.org/10.1073/pnas.0510423103
  29. Roman-Gomez J, Jimenez-Velasco A, Castillejo JA, et al (2004). The suppressor of cytokine signaling-1 is constitutively expressed in chronic myeloid leukemia and correlates with poor cytogenetic response to interferon-alpha. Haematologica, 89, 42-8.
  30. Rottapel R, Ilangumaran S, Neale C, et al (2002). The tumor suppressor activity of SOCS-1. Oncogene, 21, 4351-62. https://doi.org/10.1038/sj.onc.1205537
  31. Saelee P, Chuensumran U, Wongkham S, et al (2012). Hypermethylation of suppressor of cytokine signaling 1 in hepatocellular carcinoma patients. Asian Pac J Cancer Prev, 13, 3489-93. https://doi.org/10.7314/APJCP.2012.13.7.3489
  32. Sawyers CL (1999). Chronic myeloid leukemia. N Engl J Med, 340, 1330-40. https://doi.org/10.1056/NEJM199904293401706
  33. Shah NP, Nicoll JM, Nagar B, et al (2002). Multiple BCR-ABL kinase domain mutations confer polyclonal resistance to the tyrosine kinase inhibitor imatinib (STI571) in chronic phase and blast crisis chronic myeloid leukemia. Cancer cell, 2, 117-25. https://doi.org/10.1016/S1535-6108(02)00096-X
  34. Smith CC, Shah NP (2012). Mechanisms of Resistance to Targeted Therapies in Acute Myeloid Leukemia and Chronic Myeloid Leukemia. Am Soc Clin Oncol Educ Book, 10, 1092-9118.
  35. Sutherland KD, Lindeman GJ, Choong DY, et al (2004). Differential hypermethylation of SOCS genes in ovarian and breast carcinomas. Oncogene, 23, 7726-33. https://doi.org/10.1038/sj.onc.1207787
  36. Tamiya T, Kashiwagi I, Takahashi R, Yasukawa H, Yoshimura A (2011). Suppressors of cytokine signaling (SOCS) proteins and JAK/STAT pathways: regulation of T-cell inflammation by SOCS1 and SOCS3. Arterioscler Thromb Vasc Biol, 31, 980-5. https://doi.org/10.1161/ATVBAHA.110.207464
  37. Villuendas R, Steegmann JL, Pollan M, et al (2006). Identification of genes involved in imatinib resistance in CML: a gene-expression profiling approach. Leukemia, 20, 1047-54. https://doi.org/10.1038/sj.leu.2404197
  38. Yoshikawa H, Matsubara K, Qian GS, et al (2001). SOCS-1, a negative regulator of the JAK/STAT pathway, is silenced by methylation in human hepatocellular carcinoma and shows growth-suppression activity. Nat Genet, 28, 29-35.
  39. Zhou J, Bi C, Janakakumara JV, et al (2009). Enhanced activation of STAT pathways and overexpression of survivin confer resistance to FLT3 inhibitors and could be therapeutic targets in AML. Blood, 113, 4052-62. https://doi.org/10.1182/blood-2008-05-156422

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