Abstract
Bendamustine combined with other drugs is clinically efficacious for some adult lymphoid malignancies, but to date there are no reports of the use of such combinatorial approaches in pediatric patients. We investigated the in vitro activity of bendamustine combined with other antimetabolite drugs on B cell precursor acute lymphoblastic leukemia (BCP-ALL) cell lines established from pediatric patients with refractory or relapsed ALL. We also developed a mathematically drown improved isobologram method to assess the data objectively. Three BCP-ALL cell lines; YCUB-2, YCUB-5, and YCUB-6, were simultaneously exposed to various concentrations of bendamustine and cladribine, cytarabine, fludarabine, or clofarabine. Cell growth inhibition was determined using the WST-8 assay. Combinatorial effects were estimated using our improved isobologram method with IC80 (drug concentration corresponding to 80 % of maximum inhibition). Bendamustine alone inhibited ALL cell growth with mean IC80 values of 11.30–18.90 μg/ml. Combinations of bendamustine with other drugs produced the following effects: (1) cladribine; synergistic-to-additive on all cell lines; (2) cytarabine; synergistic-to-additive on YCUB-5 and YCUB-6, and synergistic-to-antagonistic on YCUB-2; (3) fludarabine; additive-to-antagonistic on YCUB-5, and synergistic-to-antagonistic on YCUB-2 and YCUB-6; (4) clofarabine; additive-to-antagonistic on all cell lines. Flow cytometric analysis also showed the combination effects of bendamustine and cladribine. Bendamustine/cladribine or bendamustine/cytarabine may thus represent a promising combination for salvage treatment in childhood ALL.
Similar content being viewed by others
References
Tageja N, Nagi J. Bendamustine: something old, something new. Cancer Chemother Pharmacol. 2010;66:413–23.
Glode AE, Jarkowski A. Bendamustine: a new treatment option for chronic lymphocytic leukemia. Pharmacotherapy. 2009;29:1375–84.
Hagemeister F, Manoukian G. Bendamustine in the treatment of non-Hodgkin’s lymphomas. Onco Targets Ther. 2009;2:269–79.
Gil L, Kazmierczak M, Kroll-Balcerzak R, Komarnicki M. Bendamustine-based therapy as first-line treatment for non-Hodgkin lymphoma. Med Oncol. 2014;31:944.
Fraser C, Brown P, Megason G, Ahn HS, Cho B, Kirov I, et al. Open-label bendamustine monotherapy for pediatric patients with relapsed or refractory acute leukemia: efficacy and tolerability: safety and Efficacy Outcomes. J Pediatr Hematol Oncol. 2014;36:e212–8.
Tageja N. Bendamustine: safety and efficacy in the management of indolent non-hodgkins lymphoma. Clin Med Insights Oncol. 2011;5:145–56.
Khouri IF, Wei W, Korbling M, Turturro F, Ahmed S, Alousi A, et al. BFR (bendamustine, fludarabine, and rituximab) allogeneic conditioning for chronic lymphocytic leukemia/lymphoma: reduced myelosuppression and GVHD. Blood. 2014;124:2306–12.
Schwänen C, Hecker T, Hübinger G, Wölfle M, Rittgen W, Bergmann L, et al. In vitro evaluation of bendamustine induced apoptosis in B-chronic lymphocytic leukemia. Leukemia. 2002;16:2096–105.
Chow KU, Boehrer S, Napieralski S, Nowak D, Knau A, Hoelzer D, et al. In AML cell lines Ara-C combined with purine analogues is able to exert synergistic as well as antagonistic effects on proliferation, apoptosis and disruption of mitochondrial membrane potential. Leuk Lymphoma. 2003;44:165–73.
Visco C, Castegnaro S, Chieregato K, Bernardi M, Albiero E, Zanon C, et al. The cytotoxic effects of bendamustine in combination with cytarabine in mantle cell lymphoma cell lines. Blood Cells Mol Dis. 2012;48:68–75.
Chow KU, Boehrer S, Geduldig K, Krapohl A, Hoelzer D, Mitrou PS, et al. In vitro induction of apoptosis of neoplastic cells in low-grade non-Hodgkin’s lymphomas using combinations of established cytotoxic drugs with bendamustine. Haematologica. 2001;86:485–93.
Hijiya N, Thomson B, Isakoff MS, Silverman LB, Steinherz PG, Borowitz MJ, et al. Phase 2 trial of clofarabine in combination with etoposide and cyclophosphamide in pediatric patients with refractory or relapsed acute lymphoblastic leukemia. Blood. 2011;118:6043–9.
Shukla N, Kobos R, Renaud T, Steinherz LJ, Steinherz PG. Phase II trial of clofarabine with topotecan, vinorelbine, and thiotepa in pediatric patients with relapsed or refractory acute leukemia. Pediatr Blood Cancer. 2014;61:431–5.
Steel GG, Peckham MJ. Exploitable mechanisms in combined radiotherapy-chemotherapy: the concept of additivity. Int J Radiat Oncol Biol Phys. 1979;5:85–91.
Chou TC, Talaly P. A simple generalized equation for the analysis of multiple inhibitions of Michaelis-Menten kinetic systems. J Biol Chem. 1977;252:6438–42.
Chou TC, Talalay P. Quantitative analysis of dose-effect relationships: the combined effects of multiple drugs or enzyme inhibitors. Adv Enzyme Regul. 1984;22:27–55.
Chou TC. Theoretical basis, experimental design, and computerized simulation of synergism and antagonism in drug combination studies. Pharmacol Rev. 2006;58:621–81.
Chou TC. Preclinical versus clinical drug combination studies. Leuk Lymphoma. 2008;49:2059–80.
Chou TC. Drug combination studies and their synergy quantification using the Chou-Talalay method. Cancer Res. 2010;70:440–6.
Goto H, Naruto T, Tanoshima R, Kato H, Yokosuka T, Yanagimachi M, et al. Chemo-sensitivity in a panel of B-cell precursor acute lymphoblastic leukemia cell lines, YCUB series, derived from children. Leuk Res. 2009;33:1386–91.
Kano Y, Ohnuma T, Okano T, Holland JF. Effects of vincristine in combination with methotrexate and other antitumor agents in human acute lymphoblastic leukemia cells in culture. Cancer Res. 1988;48:351–6.
Kano Y, Suzuki K, Akutsu M, Suda K, Inoue Y, Yoshida M, et al. Effects of CPT-11 in combination with other anti-cancer agents in culture. Int J Cancer. 1992;50:604–10.
Kano Y, Akutsu M, Tsunoda S, Mori K, Suzuki K, Adachi KI. In vitro schedule-dependent interaction between paclitaxel and SN-38 (the active metabolite of irinotecan) in human carcinoma cell lines. Cancer Chemother Pharmacol. 1998;42:91–8.
Kano Y, Akutsu M, Tsunoda S, Mano H, Sato Y, Honma Y, et al. In vitro cytotoxic effects of a tyrosine kinase inhibitor STI571 in combination with commonly used antileukemic agents. Blood. 2001;97:1999–2007.
Chacar C, Jabbour E, Ravandi F, Borthakur G, Kadia T, Estrov Z, et al. Phase I-II study of bendamustine in patients with acute leukemia and high risk myelodysplastic syndrome. Clin Lymphoma Myeloma Leuk. 2012;12:197–200.
Fichtner I, Arndt D, Reszka R. Antineoplastic activity and toxicity of some alkylating cytostatics (cyclophosphamide, CCNU, cytostasan) encapsulated in liposomes in different murine tumour models. J Microencapsul. 1986;3:77–87.
Carson DA, Wasson DB, Taetle R, et al. Specific toxicity of 2-chlorodeoxyadenosine toward resting and proliferating human lymphocytes. Blood. 1983;62:737–43.
Visco C, Castegnaro S, Chieregato K, Bernardi M, Albiero E, Zanon C, et al. The cytotoxic effects of bendamustine in combination with cytarabine in mantle cell lymphoma cell lines. Blood Cells Mol Dis. 2012;48:68–75.
Castegnaro S, Visco C, Chieregato K, Bernardi M, Albiero E, Zanon C, et al. Cytosine arabinoside potentiates the apoptotic effect of bendamustine on several B- and T-cell leukemia/lymphoma cells and cell lines. Leuk Lymphoma. 2012;53:2262–8.
Author information
Authors and Affiliations
Corresponding author
Ethics declarations
Conflict of interest
The authors declare that they have no conflict of interest.
About this article
Cite this article
Goto, S., Goto, H. & Yokosuka, T. The combination effects of bendamustine with antimetabolites against childhood acute lymphoblastic leukemia cells. Int J Hematol 103, 572–583 (2016). https://doi.org/10.1007/s12185-016-1952-z
Received:
Revised:
Accepted:
Published:
Issue Date:
DOI: https://doi.org/10.1007/s12185-016-1952-z