The Effect of Ursodeoxycholic acid and N-acetyl cysteine on Lymphoblast Viability
,
Abstract
Aim: To investigate invitro ursodeoxy cholic acid (UDCA) and N-acetyl cystein (NAC) effect on blast cell viability in children newly diagnosed with acute lymphoblastic leukemia (ALL). Patients and Methods: Samples were obtained from a total of 52 newly diagnosed ALL patients aged 1 to 17 years. UDCA and NAC were added at clinical relevant concentrations (0-300 micrograms) onto 5x10^5 cells were treated at room temperature in dark place. Untreated and treated cells were stained with 7AAD PE and analyzed by flow cytometry. Results: Median (IQR) blast percentage and incubation time were 90% (11) and 18 (1.5) hours, respectively. Dead/live blast cells ratio (7AAD+) was lower in lymphoblasts treated with all NAC concentration than untreated controls (p < 0.001). The use of NAC was noted to, regardless of concentration, contributed to lymphoblasts viability. On the contrary, dead/live blast cells ratio in samples treated with UDCA at the above-mentioned concentrations was relatively high, suggesting the protection role for both hepatotoxicty and against leukemia. However, the difference was not statistically significant (P >0.05). There was also no correlation between different doses of UCDA and NAC regarding blast cell viability (P > 0.232). Conclusion: The present study showed that in vitro NAC use had a protective effect on lymphoblast viability in newly diagnosed ALL patients before start of chemotherapy, and also patient-derived ALL cells can be successfully analyzed ex vivo in a short and different period of time without loss of blasts.
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9. Dworzak MN, Gaipa G, Ratei R, et al. Standardization of flow cytometric minimal residual disease evaluation in acute lymphoblastic leukemia: Multicentric assessment is feasible. Cytometry B Clin Cytom 2008;74:331-340.
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11. Zembruski NC, Stache V, Haefeli WE, et al. 7-Aminoactinomycin D for apoptosis staining in flow cytometry. Anal Biochem. 2012;429:79-81.
12. Ratei R, Basso G, Dworzak M, et al, Monitoring treatment response of childhood precursor B-cell acute lymphoblastic leukemia in the AIEOP-BFM-ALL 2000 protocol with multiparameter flow cytometry: predictive impact of early blast reduction on the remission status after induction. Leukemia 2009; 23:528-534.
13. Philpott NJ, Turner AJ, Scopes J, et al. The use of 7-amino actinomycin D in identifying apoptosis: simplicity of use and broad spectrum of application compared with other techniques. Blood 1996;87:2244-2251.
14. Stacchini A, Aliberti S, Demurtas A, et al. Flow cytometry identification of nonhemopoietic neoplasms during routine immunophenotyping. Int J Lab Hematol 2019;41:208-217.
15. Ayuk FA, Atassi N, Schuch G, et al. Complement-dependent and complement-independent cytotoxicity of polyclonal antithymocyte globulins in chronic lymphocytic leukemia. Leuk Res 2008;32:1200-1206.
16. Bordbar M, Shakibazad N, Fattahi M, et al. Effect of ursodeoxycholic acid and vitamin E in the prevention of liver injury from methotrexate in pediatric leukemia. Turk J Gastroenterol 2018;29:203-209.
17. Kocyigit A, Guler EM, Karatas E, et al. Dose-dependent proliferative and cytotoxic effects of melatonin on human epidermoid carcinoma and normal skin fibroblast cells. Mutat Res Genet Toxicol Environ Mutagen 2018;829-830:50-60.
18. Uraz S, Tahan V, Aygun C, et al. Role of ursodeoxycholic acid in prevention of methotrexate-induced liver toxicity. Dig Dis Sci 2008;53:1071–1077.
19. Stary J, Zimmermann M, Campbell M, et al. Intensive chemotherapy for childhood acute lymphoblastic leukemia: results of the randomized intercontinental trial ALL IC-BFM 2002. J Clin Oncol 2014;32:174-184.
20. Schoeneberger H, Belz K, Schenk B, et al. Impairment of antioxidant defense via glutathione depletion sensitizes acute lymphoblastic leukemia cells for Smac mimetic-induced cell death.Oncogene 2015;34:4032-4043.
21. Uckun FM1, Stork L, Seibel N, et al. Residual bone marrow leukemic progenitor cell burden after induction chemotherapy in pediatric patients with acute lymphoblastic leukemia. Clin Cancer Res 2000;6:3123-3130.
22. Parikh SK, Uparkar UP. Assessment of minimal residual disease in childhood acute lymphoblastic leukemia. J Appl Hematol 2016;7:47-53.
23. Kong Y, Song Y, Tang FF et al. N-acetyl-L-cysteine improves mesenchymal stem cell function in prolonged isolated thrombocytopenia post-allotransplant. Br J Haematol 2018;180:863-878.
24. Dworzak MN, Fröschl G, Printz D, et al. Prognostic significance and modalities of flow cytometric minimal residual disease detection in childhood acute lymphoblastic leukemia. Blood 2002;99:1952-1958.
25. Kong Y, Shi MM, Zhang YY, et al. N-acetyl-L-cysteine improves bone marrow endothelial progenitor cells in prolonged isolated thrombocytopenia patients post allogeneic hematopoietic stem cell transplantation. Am J Hematol 2018;93:931-942.
26. Al-Rubeai M, Welzenbach K, Lloyd DR, et al. A rapid method for evaluation of cell number and viability by flow cytometry. Cytotechnology 1997;24:161-168.
2. Ebbesen MS, Nygaard U, Rosthøj S, et al. Hepatotoxicity During Maintenance Therapy and Prognosis in Children With Acute Lymphoblastic Leukemia. J Pediatr Hematol Oncol 2017; 39:161- 166.
3. Okamura A, Nishimura M, Sanada Y, et al. L-Asparaginase-induced fulminating liver dysfunction. Int J Hematol 2013;98:6-7.
4. King PD, Perry MC. Hepatotoxicity of chemotherapy. Oncologist. 2001;6:162–176.
5. Giordano C, Rivas J, Zervos X. An Update on Treatment of Drug-Induced Liver Injury. J Clin Transl Hepatol 2014;2:74-79.
6. Al-Tonbary Y, Al-Haggar M, El-Ashry R, et al. Vitamin e and N-acetylcysteine as antioxidant adjuvant therapy in children with acute lymphoblastic leukemia. Adv Hematol 2009;689639.
7. Pal D, Blair HJ, Elder A, et al. Long-term in vitro maintenance of clonal abundance and leukaemia-initiating potential in acute lymphoblastic leukaemia. Leukemia 2016;30:1691–700.
8. Modvig S, Madsen HO, Siitonen SM, Rosthøj S, Tierens A, Juvonen V, et al. Minimal residual disease quantification by flow cytometry provides reliable risk stratification in T-cell acute lymphoblastic leukemia. Leukemia 2019;33:1324-1336.
9. Dworzak MN, Gaipa G, Ratei R, et al. Standardization of flow cytometric minimal residual disease evaluation in acute lymphoblastic leukemia: Multicentric assessment is feasible. Cytometry B Clin Cytom 2008;74:331-340.
10. Diks AM, Bonroy C, Teodosio C, et al. Impact of blood storage and sample handling on quality of high dimensional flow cytometric data in multicenter clinical research. J Immunol Methods 2019;475:112616.
11. Zembruski NC, Stache V, Haefeli WE, et al. 7-Aminoactinomycin D for apoptosis staining in flow cytometry. Anal Biochem. 2012;429:79-81.
12. Ratei R, Basso G, Dworzak M, et al, Monitoring treatment response of childhood precursor B-cell acute lymphoblastic leukemia in the AIEOP-BFM-ALL 2000 protocol with multiparameter flow cytometry: predictive impact of early blast reduction on the remission status after induction. Leukemia 2009; 23:528-534.
13. Philpott NJ, Turner AJ, Scopes J, et al. The use of 7-amino actinomycin D in identifying apoptosis: simplicity of use and broad spectrum of application compared with other techniques. Blood 1996;87:2244-2251.
14. Stacchini A, Aliberti S, Demurtas A, et al. Flow cytometry identification of nonhemopoietic neoplasms during routine immunophenotyping. Int J Lab Hematol 2019;41:208-217.
15. Ayuk FA, Atassi N, Schuch G, et al. Complement-dependent and complement-independent cytotoxicity of polyclonal antithymocyte globulins in chronic lymphocytic leukemia. Leuk Res 2008;32:1200-1206.
16. Bordbar M, Shakibazad N, Fattahi M, et al. Effect of ursodeoxycholic acid and vitamin E in the prevention of liver injury from methotrexate in pediatric leukemia. Turk J Gastroenterol 2018;29:203-209.
17. Kocyigit A, Guler EM, Karatas E, et al. Dose-dependent proliferative and cytotoxic effects of melatonin on human epidermoid carcinoma and normal skin fibroblast cells. Mutat Res Genet Toxicol Environ Mutagen 2018;829-830:50-60.
18. Uraz S, Tahan V, Aygun C, et al. Role of ursodeoxycholic acid in prevention of methotrexate-induced liver toxicity. Dig Dis Sci 2008;53:1071–1077.
19. Stary J, Zimmermann M, Campbell M, et al. Intensive chemotherapy for childhood acute lymphoblastic leukemia: results of the randomized intercontinental trial ALL IC-BFM 2002. J Clin Oncol 2014;32:174-184.
20. Schoeneberger H, Belz K, Schenk B, et al. Impairment of antioxidant defense via glutathione depletion sensitizes acute lymphoblastic leukemia cells for Smac mimetic-induced cell death.Oncogene 2015;34:4032-4043.
21. Uckun FM1, Stork L, Seibel N, et al. Residual bone marrow leukemic progenitor cell burden after induction chemotherapy in pediatric patients with acute lymphoblastic leukemia. Clin Cancer Res 2000;6:3123-3130.
22. Parikh SK, Uparkar UP. Assessment of minimal residual disease in childhood acute lymphoblastic leukemia. J Appl Hematol 2016;7:47-53.
23. Kong Y, Song Y, Tang FF et al. N-acetyl-L-cysteine improves mesenchymal stem cell function in prolonged isolated thrombocytopenia post-allotransplant. Br J Haematol 2018;180:863-878.
24. Dworzak MN, Fröschl G, Printz D, et al. Prognostic significance and modalities of flow cytometric minimal residual disease detection in childhood acute lymphoblastic leukemia. Blood 2002;99:1952-1958.
25. Kong Y, Shi MM, Zhang YY, et al. N-acetyl-L-cysteine improves bone marrow endothelial progenitor cells in prolonged isolated thrombocytopenia patients post allogeneic hematopoietic stem cell transplantation. Am J Hematol 2018;93:931-942.
26. Al-Rubeai M, Welzenbach K, Lloyd DR, et al. A rapid method for evaluation of cell number and viability by flow cytometry. Cytotechnology 1997;24:161-168.
| Files | ||
| Issue | Vol 13 No 1 (2021) | |
| Section | Original Articles | |
| DOI | https://doi.org/10.18502/bccr.v13i1.8829 | |
| Keywords | ||
| children acute lymphoblastic leukemia ursodeoxy cholic acid N-acetyl cystein | ||
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How to Cite
1.
Aycicek A, Tahtakesen TN, Bayram C. The Effect of Ursodeoxycholic acid and N-acetyl cysteine on Lymphoblast Viability. Basic Clin Cancer Res. 2022;13(1):55-62.
