Arsenic trioxide and ascorbic acid interfere with the BCL2 family genes in patients with myelodysplastic syndromes: an ex-vivo study

Journal of Hematology & Oncology

Alessandro Levis¹  Mario Petrini⁴  Daniela Cilloni⁶  Giuseppe A Palumbo²  Emanuela Messa¹  Daniela Gioia¹  Iacopo Petrini⁵  Flavia Salvi¹  Francesca Guerrini⁴  Sara Galimberti³ 

[1] S.O.C. di Ematologia, Azienda Ospedaliera SS Antonio e Biagio, Alessandria, Italy;Department of Clinical and Molecular Bio-Medicine, Section of Hematology, Oncology and General Pathology, Catania, Italy;Division of Haematology, Department of Oncology, Transplant and Advances in Medicine, University of Pisa, Via Roma 67, Pisa, 56126, Italy;Department of Oncology, Transplant, New Advances in Medicine, Section of Hematology, University of Pisa, Pisa, Italy;Department of Oncology, Transplant and New Advances in Medicine, BIOS, University of Pisa, Pisa, Italy;Department of Clinical and Biological Sciences, University of Turin, Turin, Italy

关键词: Apoptosis;    MDS;    Myelodysplastic syndromes;    Ascorbic acid;    ATO;   

Others  :  822186
DOI  :  10.1186/1756-8722-5-53

 received in 2012-07-09, accepted in 2012-08-27,  发布年份 2012

【 摘 要 】

Background

Arsenic Trioxide (ATO) is effective in about 20% of patients with myelodysplasia (MDS); its mechanisms of action have already been evaluated in vitro, but the in vivo activity is still not fully understood. Since ATO induces apoptosis in in vitro models, we compared the expression of 93 apoptotic genes in patients’ bone marrow before and after ATO treatment. For this analysis, we selected 12 patients affected by MDS who received ATO in combination with Ascorbic Acid in the context of the Italian clinical trial NCT00803530, EudracT Number 2005-001321-28.

Methods

Real-time PCR quantitative assays for genes involved in apoptosis were performed using TaqMan® Assays in 384-Well Microfluidic Cards “TaqMan® Human Apoptosis Array”.

Quantitative RT-PCR for expression of EVI1 and WT1 genes was also performed. Gene expression values (Ct) were normalized to the median expression of 3 housekeeping genes present in the card (18S, ACTB and GAPDH).

Results

ATO treatment induced up-regulation of some pro-apoptotic genes, such as HRK, BAK1, CASPASE-5, BAD, TNFRSF1A, and BCL2L14 and down-regulation of ICEBERG. In the majority of cases with stable disease, apoptotic gene expression profile did not change, whereas in cases with advanced MDS more frequently pro-apoptotic genes were up-regulated. Two patients achieved a major response: in the patient with refractory anemia the treatment down-regulated 69% of the pro-apoptotic genes, whereas 91% of the pro-apoptotic genes were up-regulated in the patient affected by refractory anemia with excess of blasts-1. Responsive patients showed a higher induction of BAD than those with stable disease. Finally, WT1 gene expression was down-regulated by the treatment in responsive cases.

Conclusions

These results represent the basis for a possible association of ATO with other biological compounds able to modify the apoptotic pathways, such as inhibitors of the BCL2 family.

【 授权许可】

   
2012 Galimberti et al.; licensee BioMed Central Ltd.

【 图 表 】

Figure 1.

Figure 2.

Figure 3.

Figure 4.

【 参考文献 】

• [1]Ho SY, Wu WJ, Chiu HW, Chen YA, Ho YS, Guo HR, Wang YJ: Arsenic trioxide and radiation enhance apoptotic effects in HL-60 cells through increased ROS generation and regulation of JNK and p38 MAPK signaling pathways. Chem Biol Interact 2011, 193:162-167.
• [2]Schiller GJ, Slack J, Hainsworth JD, Mason J, Saleh M, Rizzieri D, Douer D, List AF: Phase II multicenter study of arsenic trioxide in patients with myelodysplastic syndromes. J Clin Oncol 2006, 24:2456-2464.
• [3]Vey N, Bosly A, Guerci A, Feremans W, Dombret H, Dreyfus F, Bowen D, Burnett A, Dennis M, Ribrag V, Casadevall N, Legros L, Fenaux P: Arsenic trioxide in patients with myelodysplastic syndromes: a phase II multicenter study. J Clin Oncol 2006, 24:2465-2471.
• [4]Zheng WL, Zhang GS, Xu YX, Shen JK, Dai CW, Pei MF: Arsenic trioxide, thalidomide and retinoid acid combination therapy in higher risk myelodysplastic syndrome patients. Leuk Res 2008, 32:251-254.
• [5]Perkins C, Kim CN, Fang G, Bhalla KN: Arsenic induces apoptosis of multidrug-resistant human myeloid leukemia cells that express Bcr-Abl or overexpress MDR, MRP, Bcl-2, or Bcl-x(L). Blood 2000, 95:1014-1022.
• [6]Scholz C, Richter A, Lehmann M, Schulze-Osthoff K, Dorken B, Daniel PT: Arsenic trioxide induces regulated, death receptor-independent cell death through a Bcl-2-controlled pathway. Oncogene 2005, 24:7031-7042.
• [7]Mathas S, Lietz A, Janz M, Hinz M, Jundt F, Scheidereit C, Bommert K, Dorken B: Inhibition of NF-kappa B essentially contributes to arsenic-induced apoptosis. Blood 2003, 102:1028-1034.
• [8]Yedjou C, Tchounwou P, Jenkins J, McMurray R: Basic mechanisms of arsenic trioxide (ATO)-induced apoptosis in human leukemia (HL-60) cells. J Hematol Oncol 2010, 23:28.
• [9]Huang XJ, Wiernik PH, Klein RS, Gallagher RE: Arsenic trioxide induces apoptosis of myeloid leukemia cells by activation of caspases. Med Oncol 1999, 16:58-64.
• [10]Roboz GJ, Dias S, Lam G, Lane WJ, Soignet SL, Warrell RP Jr, Rafii S: Arsenic trioxide induces dose- and time-dependent apoptosis of endothelium and may exert an antileukemic effect via inhibition of angiogenesis. Blood 2000, 96:1525-1530.
• [11]Canestraro M, Galimberti S, Savli H, Palumbo GA, Tibullo D, Nagy B, Guerrini F, Piaggi S, Cine N, Metelli MR, Petrini M: Synergistic antiproliferative effect of arsenic trioxide combined with bortezomib in HL60 cell line and primary blasts from patients affected by myeloproliferative disorders. Cancer Genet Cytogenet 2010, 199:110-120.
• [12]Galimberti S, Ghio F, Guerrini F, Ciabatti E, Grassi S, Ferreri MI, Petrini M: WT1 expression levels at diagnosis could predict long-term time-to-progression in adult patients affected by acute myeloid leukaemia and myelodysplastic syndromes. Br J Haematol 2010, 149:451-454.
• [13]Shackelford D, Kenific C, Blusztajn A, Waxman S, Ren R: Targeted degradation of the AML1/MDS1/EVI1 oncoprotein by arsenic trioxide. Cancer Res 2006, 66:11360-11369.
• [14]Malcovati L, Germing U, Kuendgen A, Della Porta MG, Pascutto C, Invernizzi R, Giagounidis A, Hildebrandt B, Bernasconi P, Knipp S, Strupp C, Lazzarino M, Aul C, Cazzola M: Time-dependent prognostic scoring system for predicting survival and leukemic evolution in myelodysplastic syndromes. J Clin Oncol 2007, 25:3503-3510.
• [15]Sanz GF, Sanz MA, Greenberg PL: Prognostic factors and scoring systems in myelodysplastic syndromes. Haematologica 1998, 83:358-368.
• [16]Cheson BD, Greenberg PL, Bennett JM, Lowenberg B, Wijermans PW, Nimer SD, Pinto A, Beran M, de Witte TM, Stone RM, Mittelman M, Sanz GF, Gore SD, Schiffer CA, Kantarjian H: Clinical application and proposal for modification of the International Working Group (IWG) response criteria in myelodysplasia. Blood 2006, 108:419-425.
• [17]Boudard D, Sordet O, Vasselon C, Revol V, Berthéas MF, Freyssenet D, Viallet A, Piselli S, Guyotat D, Campos L: Expression and activity of caspases 1 and 3 in myelodysplastic syndromes. Leukemia 2000, 14:2045-2051.
• [18]Navas T, Zhou L, Estes M, Haghnazari E, Nguyen AN, Mo Y, Pahanish P, Mohindru M, Cao T, Higgins LS, Platanias LC, List A, Verma A, Bhagat T, Gajavelli S, Kambhampati S: Inhibition of p38alpha MAPK disrupts the pathological loop of proinflammatory factor production in the myelodysplastic syndrome bone marrow microenvironment. Leuk Lymphoma 2008, 49:1963-1975.
• [19]Schall TJ, Lewis M, Koller KJ, Lee A, Rice GC, Wong GH, Gatanaga T, Granger GA, Lentz R, Raab H: Molecular cloning and expression of a receptor for human tumor necrosis factor. Cell 1990, 61:361-370.
• [20]Nolte F, Hofmann WK: Molecular mechanisms involved in the progression of myelodysplastic syndrome. Future Oncol 2010, 6:445-455.
• [21]Keilholz U, Letsch A, Busse A, Asemissen AM, Bauer S, Blau IW, Hofmann WK, Uharek L, Thiel E, Scheibenbogen C: A clinical and immunologic phase 2 trial of Wilms tumor gene product 1 (WT1) peptide vaccination in patients with AML and MDS. Blood 2009, 113:6541-6548.
• [22]Tse C, Shoemaker AR, Adickes J, Anderson MG, Chen J, Jin S, Johnson EF, Marsh KC, Mitten MJ, Nimmer P, Roberts L, Tahir SK, Xiao Y, Yang X, Zhang H, Fesik S, Rosenberg SH, Elmore SW: ABT-263: a potent and orally bioavailable Bcl-2 family inhibitor. Cancer Res 2008, 68:3421-3428.