In vitro drug sensitivity in canine lymphoma

Open access


Introduction: Due to the high heterogeneity of canine lymphoma, the aim of the present study was to test in vitro the chemosensitivity of canine high-grade primary lymphoma cells to various cytostatic drugs commonly used to treat dogs: 4-HO-cyclophosphamide, doxorubicin, dexamethasone, prednisolone, vincristine, etoposide, chlorambucil, lomustine, and cytosine arabinoside. Material and Methods: To determine the cell viability and drug ability to induce apoptosis two different tests were used: an MTT assay and annexin V/propidium iodide staining. Results: Both in vitro tests were found to be useful tools. Significant differences in the sensitivity, depending on the drug type, between B-, T- and mixed/null-type lymphoma cells were found for the majority of the tested drugs. B-type cells were the most sensitive in vitro, whereas T-type cells seemed to be the most resistant. Doxorubicin, chlorambucil, etoposide, and vincristine most strongly reduced the cell viability and induced apoptosis. Conclusion: In vitro assays, such as the MTT test and especially the annexin V/PI assay, may be useful tools for predicting a response to the treatment of high-grade lymphoma in dogs or improving the treatment outcomes in individual animals.


  • 1. Den Boer M.L., Harms D.O., Pieters R., Kazemier K.M., Gobel U., Körholz D., Graubner U., Haas R.J., Jorch N., Spaar H.J., Kaspers G.J., Kamps W.A., Van der Does-Van den Berg A., Van Wering E.R., Veerman A.J., Janka-Schaub G.E.: Patient stratification based on prednisolone-vincristineasparaginase resistance profiles in children with acute lymphoblastic leukemia. J Clin Oncol 2003, 21, 262-3268.

  • 2. Escherich G., Tröger A., Göbel U., Graubner U., Pekrun A., Jorch N., Kaspers G., Zimmermann M., zur Stadt U., Kazemier K., Pieters R., Den Boer M.L., Horstmann M., Janka G.E.: The long-term impact of in vitro drug sensitivity on risk stratification and treatment outcome in acute lymphoblastic leukemia of childhood (CoALL 06-97). Haematologica 2011, 96, 854-862.

  • 3. Fedele C., Tothill R.W., McArthur G.A.: Navigating the challenge of tumor heterogeneity in cancer therapy. Cancer Discov 2014, 4,146-148.

  • 4. Gaurnier-Hausser A., Patel R., Baldwin A.S., May M.J., Mason N.J.: NEMO-binding domain peptide inhibits constitutive NF-kappaB activity and reduces tumor burden in a canine model of relapsed, refractory diffuse large B-cell lymphoma. Clin Cancer Res 2011, 17, 4661-4671.

  • 5. Hatok J., Babusikova E., Matakova T., Mistuna D., Dobrota D., Racay P.: In vitro assays for the evaluation of drug resistance in tumor cells. Clin Exp Med 2009, 9, 1-7.

  • 6. Ito D., Frantz A.M., Modiano J.F.: Canine lymphoma as a comparative model for human non-Hodgkin lymphoma: recent progress and applications. Vet Immunol Immunopathol 2014, 24, 16.

  • 7. Ito D., Frantz A.M., Williams C., Thomas R., Burnett R.C., Avery A.C., Modiano J.F.: CD40 ligand is necessary and sufficient to support primary diffuse large B-cell lymphoma cells in culture: a tool for in vitro preclinical studies with primary B-cell malignancies. Leuk Lymphoma 2012, 53, 1390-1398.

  • 8. Kaspers G.J., Pieters R., Van Zantwijk C.H., De Laat P.A., De Waal F.C., Van Wering E.R., Veerman A.J.: In vitro drug sensitivity of normal peripheral blood lymphocytes and childhood leukaemic cells from bone marrow and peripheral blood. Br J Cancer 1991, 64, 469-474.

  • 9. Kitada S., Andersen J., Akar S., Zapata J.M., Takayama S., Krajewski S., Wang H.G., Zhang X., Bullrich F., Croce C.M., Rai K., Hines J., Reed J.C.: Expression of apoptosis-regulating proteins in chronic lymphocytic leukemia: correlations with in vitro and in vivo chemoresponses. Blood 1998, 91, 3379-3389.

  • 10. Marconato L.: The staging and treatment of multicentric highgrade lymphoma in dogs: a review of recent developments and future prospects. Vet J 2011, 188, 34-38.

  • 11. Marconato L., Gelain M.E., Comazzi S.: The dog as a possible animal model for human non-Hodgkin lymphoma: a review. Hematol Oncol 2013, 31, 1-9.

  • 12. Matsuda A., Tanaka A., Muto S., Ohmori K., Furusaka T., Jung K., Karasawa K., Okamoto N., Oida K., Itai A., Matsuda H.: A novel NF-kappaB inhibitor improves glucocorticoid sensitivity of canine neoplastic lymphoid cells by up-regulating expression of glucocorticoid receptors. Res Vet Sci 2010, 89, 378-382.

  • 13. Mosmann T.: Rapid colorimetric assay for cellular growth and survival: application to proliferation and cytotoxicity assays. J Immunol Methods 1983, 65, 55-63.

  • 14. Pawlak A., Rapak A., Drynda A., Poradowski D., Zbyryt I., Dzimira S., Suchański J., Obmińska-Mrukowicz B.: Immunophenotypic characterization of canine malignant lymphoma: a retrospective study of cases diagnosed in Poland Lower Silesia, over the period 2011-2013. Vet Comp Oncol 2014 doi: 10.1111/vco.12112.

  • 15. Pawlak A., Rapak A., Zbyryt I., Obminska-Mrukowicz B.: The effect of common antineoplastic agents on induction of apoptosis in canine lymphoma and leukemia cell lines. In Vivo 2014, 28, 843-850.

  • 16. Pieters R., Huismans D.R., Loonen A.H., Hählen K., van der Does-van den Berg A., van Wering E.R., Veerman A.J.: Relation of cellular drug resistance to long-term clinical outcome in childhood acute lymphoblastic leukaemia. Lancet 1991, 338, 399-403.

  • 17. Price G.S., Page R.L., Fischer B.M., Levine J.F., Gerig T.M.: Efficacy and toxicity of doxorubicin/cyclophosphamide maintenance therapy in dogs with multicentric lymphosarcoma. J Vet Intern Med 1991, 5, 259-262.

  • 18. Seiser E.L., Thomas R., Richards K.L., Kelley M.K., Moore P., Suter S.E., Breen M.: Reading between the lines: molecular characterization of five widely used canine lymphoid tumour cell lines. Vet Comp Oncol 2013, 11, 30-50.

  • 19. Styczynski, J., Wysocki M.: Is the in vitro drug resistance profile the strongest prognostic factor in childhood acute lymphoblastic leukemia? J Clin Oncol 2004, 5, 963-964.

  • 20. Vaughan A., Johnson J.L., Williams L.E.: Impact of chemotherapeutic dose intensity and hematologic toxicity on first remission duration in dogs with lymphoma treated with a chemoradiotherapy protocol. J Vet Intern Med 2007, 21, 1332-1339.

  • 21. Zandvliet M., Teske E., Schrickx J.A.: Multi-drug resistance in a canine lymphoid cell line due to increased P-glycoprotein expression, a potential model for drug-resistant canine lymphoma. Toxicol In Vitro 2014, 28, 1498-1506.

Journal of Veterinary Research

formerly Bulletin of the Veterinary Institute in Pulawy

Journal Information

IMPACT FACTOR Bull Vet Inst Pulawy 2016: 0.462

CiteScore 2016: 0.46

SCImago Journal Rank (SJR) 2015: 0.230
Source Normalized Impact per Paper (SNIP) 2015: 0.383


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