The effect of nilotinib plus arsenic trioxide on the proliferation and differentiation of primary leukemic cells from patients with chronic myoloid leukemia in blast crisis

Cancer Cell International

LiHong Chen¹  YaLing Chen¹  Nannan Li¹  Yan Du¹  Fei-fei Lv¹  Wei Wang¹ 

[1] Department of Hematology, Southeast Hospital Affiliated to Xiamen University (the 175th Hospital of Chinese PLA), NO.269, Zhanghua Middle Road, Zhangzhou 363000, Fujian, China

关键词: Differentiation;    Proliferation;    CML-BC;    AMN107;    Nilotinib;    Arsenic trioxide;   

Others  :  1138575
DOI  :  10.1186/s12935-015-0158-4

 received in 2014-07-10, accepted in 2015-01-05,  发布年份 2015

【 摘 要 】

Aim

To determine the effects of arsenic trioxide (ATO) and nilotinib (AMN107, Tasigna) alone or in combination on the proliferation and differentiation of primary leukemic cells from patients with chronic myeloid leukemia in the blast crisis phase (CML-BC).

Methods

Cells were isolated from the bone marrow of CML-BC patients and were treated with 1 μM ATO and 5 nM nilotinib, either alone or in combination. Cell proliferation was evaluated using a MTT assay. Cell morphology and the content of hemoglobin were examined with Wright-Giemsa staining and benzidine staining, respectively. The expression of cell surface markers was determined using flow cytometric analysis. The levels of mRNA and protein were analyzed using RT-PCR and Western blotting, respectively.

Results

ATO and nilotinib alone or in combination suppressed cell proliferation in a dose- and time-dependent pattern (P < 0.01 vs. control). Drug treatments promoted erythroid differentiation of CML-BC cells, with a decreased nuclei/cytoplasm ratio but increased hemoglobin content and glycophorin A (GPA) expression (P < 0.01 compared with control). In addition, macrophage and granulocyte lineage differentiation was also induced after drug treatment. The mRNA and protein levels of basic helix-loop-helix (bHLH) transcription factor T-cell acute lymphocytic leukemia protein 1 (TAL1) and B cell translocation gene 1 (BTG1) were both upregulated after 3 days of ATO and Nilotinib treatment.

Conclusions

Our findings indicated that ATO and nilotinib treatment alone or in combination greatly suppressed cell proliferation but promoted the differentiation of CML-BC cells towards multiple-lineages. Nilotinib alone preferentially induced erythroid differentiation while combined treatment with ATO preferentially induced macrophage and granulocyte lineage differentiation.

【 授权许可】

   
2015 Wang et al.; licensee BioMed Central.

【 图 表 】

Figure 1.

Figure 2.

Figure 3.

Figure 4.

Figure 5.

【 参考文献 】

• [1]Perrotti D, Jamieson C, Goldman J, Skorski T: Chronic myeloid leukemia: mechanisms of blastic transformation. J Clin Invest 2010, 120(7):2254-64.
• [2]Sachs RK, Johnsson K, Hahnfeldt P, Luo J, Chen A, Hlatky L: A multicellular basis for the origination of blast crisis in chronic myeloid leukemia. Cancer Res 2011, 71(8):2838-47.
• [3]Calabretta B, Perrotti D: The biology of CML blast crisis. Blood 2004, 103(11):4010-22.
• [4]Manley PW, Cowan-Jacob SW, Buchdunger E, Fabbro D, Fendrich G, Furet P, et al.: Imatinib: a selective tyrosine kinase inhibitor. Eur J Cancer. 2002, 38(Suppl 5):S19-27.
• [5]Weisberg E, Manley P, Mestan J, Cowan-Jacob S, Ray A, Griffin JD: AMN107 (nilotinib): a novel and selective inhibitor of BCR-ABL. Br J Cancer 2006, 94(12):1765-9.
• [6]le Coutre P, Ottmann OG, Giles F, Kim DW, Cortes J, Gattermann N, et al.: Nilotinib (formerly AMN107), a highly selective BCR-ABL tyrosine kinase inhibitor, is active in patients with imatinib-resistant or -intolerant accelerated-phase chronic myelogenous leukemia. Blood 2008, 111(4):1834-9.
• [7]Kantarjian HM, Giles F, Gattermann N, Bhalla K, Alimena G, Palandri F, et al.: Nilotinib (formerly AMN107), a highly selective BCR-ABL tyrosine kinase inhibitor, is effective in patients with Philadelphia chromosome-positive chronic myelogenous leukemia in chronic phase following imatinib resistance and intolerance. Blood 2007, 110(10):3540-6.
• [8]Hughes T, Saglio G, Branford S, Soverini S, Kim DW, Muller MC, et al.: Impact of baseline BCR-ABL mutations on response to nilotinib in patients with chronic myeloid leukemia in chronic phase. J Clin Oncol 2009, 27(25):4204-10.
• [9]Douer D, Tallman MS: Arsenic trioxide: new clinical experience with an old medication in hematologic malignancies. J Clin Oncol 2005, 23(10):2396-410.
• [10]Chen SJ, Zhou GB, Zhang XW, Mao JH, de The H, Chen Z: From an old remedy to a magic bullet: molecular mechanisms underlying the therapeutic effects of arsenic in fighting leukemia. Blood 2011, 117(24):6425-37.
• [11]El Eit RM, Iskandarani AN, Saliba JL, Jabbour MN, Mahfouz RA, Bitar NM, et al.: Effective targeting of chronic myeloid leukemia initiating activity with the combination of arsenic trioxide and interferon alpha. Int J Cancer 2014, 134(4):988-96.
• [12]Sawyers CL, Hochhaus A, Feldman E, Goldman JM, Miller CB, Ottmann OG, et al.: Imatinib induces hematologic and cytogenetic responses in patients with chronic myelogenous leukemia in myeloid blast crisis: results of a phase II study. Blood 2002, 99(10):3530-9.
• [13]Talpaz M, Silver RT, Druker BJ, Goldman JM, Gambacorti-Passerini C, Guilhot F, et al.: Imatinib induces durable hematologic and cytogenetic responses in patients with accelerated phase chronic myeloid leukemia: results of a phase 2 study. Blood 2002, 99(6):1928-37.
• [14]Kantarjian HM, Cortes J, O'Brien S, Giles FJ, Albitar M, Rios MB, et al.: Imatinib mesylate (STI571) therapy for Philadelphia chromosome-positive chronic myelogenous leukemia in blast phase. Blood 2002, 99(10):3547-53.
• [15]Kantarjian HM, O'Brien S, Cortes JE, Smith TL, Rios MB, Shan J, et al.: Treatment of philadelphia chromosome-positive, accelerated-phase chronic myelogenous leukemia with imatinib mesylate. Clin Cancer Res 2002, 8(7):2167-76.
• [16]Golemovic M, Verstovsek S, Giles F, Cortes J, Manshouri T, Manley PW, et al.: AMN107, a novel aminopyrimidine inhibitor of Bcr-Abl, has in vitro activity against imatinib-resistant chronic myeloid leukemia. Clin Cancer Res 2005, 11(13):4941-7.
• [17]O'Hare T, Corbin AS, Druker BJ: Targeted CML therapy: controlling drug resistance, seeking cure. Curr Opin Genet Dev 2006, 16(1):92-9.
• [18]Wang W, Li NN, Du Y, Lv FF, Lin GQ: FoxO3a and nilotinib-induced erythroid differentiation of CML-BC cells. Leuk Res 2013, 37(10):1309-14.
• [19]Shim MJ, Kim HJ, Yang SJ, Lee IS, Choi HI, Kim T: Arsenic trioxide induces apoptosis in chronic myelogenous leukemia K562 cells: possible involvement of p38 MAP kinase. J Biochem Mol Biol 2002, 35(4):377-83.
• [20]Zhu J, Okumura H, Ohtake S, Nakamura S, Nakao S: Arsenic trioxide induces apoptosis in leukemia/lymphoma cell lines via the CD95/CD95L system. Oncol Rep 2003, 10(3):705-9.
• [21]Yedjou CG, Moore P, Tchounwou PB: Dose- and time-dependent response of human leukemia (HL-60) cells to arsenic trioxide treatment. Int J Environ Res Public Health 2006, 3(2):136-40.
• [22]Miller WH Jr, Schipper HM, Lee JS, Singer J, Waxman S: Mechanisms of action of arsenic trioxide. Cancer Res 2002, 62(14):3893-903.
• [23]Mouthon MA, Bernard O, Mitjavila MT, Romeo PH, Vainchenker W, Mathieu-Mahul D: Expression of tal-1 and GATA-binding proteins during human hematopoiesis. Blood 1993, 81(3):647-55.
• [24]Brunet de la Grange P, Armstrong F, Duval V, Rouyez MC, Goardon N, Romeo PH, et al.: Low SCL/TAL1 expression reveals its major role in adult hematopoietic myeloid progenitors and stem cells. Blood 2006, 108(9):2998-3004.
• [25]Zhou RQ, Gong YP, Guo Y, Shan QQ, Yang X: The effect of knockdown of transcription factor SCL/TAL-1 gene on the erythroid differentiation in EPO-induced K562 cell line. Zhonghua Xue Ye Xue Za Zhi 2012, 33(6):453-6.
• [26]Aplan PD, Nakahara K, Orkin SH, Kirsch IR: The SCL gene product: a positive regulator of erythroid differentiation. EMBO J 1992, 11(11):4073-81.
• [27]Bakker WJ, Blazquez-Domingo M, Kolbus A, Besooyen J, Steinlein P, Beug H, et al.: FoxO3a regulates erythroid differentiation and induces BTG1, an activator of protein arginine methyl transferase 1. J Cell Biol 2004, 164(2):175-84.