Apoptotic markers in CD34-positive cells in acute leukemias

Ye.N. Parovichnikova1, Ye.Ye. Khodunova1, I.V. Galtseva1, S.М. Kulikov1, V.V. Troitskaya1, L.А. Kuzmina1, D.V. Shcheblyakov2, and V.G. Savchenko1

1 Hematology Research Center, RF Ministry of Health, Moscow, Russian Federation

2 N.F. Gamaleya Research Institution of Epidemiology and Microbiology, RF Ministry of Health, Moscow, Russian Federation


ABSTRACT

Objective. To evaluate expression of Bcl-2, Bax, p53, CD95, and ACE on CD34+ cells of peripheral blood and bone marrow during induction chemotherapy in the patients with newly diagnosed acute leukemia.

Materials and methods. Expression of Bcl-2, Bax, p53, CD95, and ACE on CD34+ cells of the peripheral blood and bone marrow in 23 patients with AL (14 AML and 9 ALL) was measured using flow cytometry analysis. Peripheral blood and bone marrow samples were analyzed before chemotherapy and during the induction course: on Days +8, +21 (blood only), and +36–38. The control group consisted of 8 healthy donors.

Results. Bcl-2 expression on CD34+ sells in BM was 34.8 ± 6 % and significantly higher compared to the donors (11.5 ± 1.8 %) at the time of diagnosis. On Days +36–38 after the onset of chemotherapy, no significant difference between the patients and control groups were found. CD34/Bax coexpression in BM cells of ALL patients was significantly higher than in AML patients and donors. ACE and p53 expression on CD34+ cells in AL patients before and during chemotherapy was significantly lower than in the donors. CD34/ACE coexpression in PB and BM cells of AL patients and donors showed no significant differences at any time-points of evaluation.

Conclusion. The above changes suggest the imbalance between the pro- and anti-apoptotic proteins in AL patients. After chemotherapy, the expression profile of these proteins considerably changed, but did not reach the healthy donor values.


Keywords: acute leukemias, apoptosis, expression of Bcl-2, Bax, р53, CD95, and ACE.

Read in  PDF (RUS)pdficon


Refernces

  1. Hengartner M.O. The biochemistry of apoptosis. Nature 2000; 407(6805): 770–6.
  2. Lodish H., Berk A., Matsudaira P. et al. Molecular cell biology, 5th edn. New York: W.H. Freeman and Company, 2003: 961.
  3. Salminen A., Ojala J., Kaarniranta K. Apoptosis and aging: increased resistance to apoptosis enhances the aging process. Cell. Mol. Life Sci. 2011; 68(6): 1021–31.
  4. Spencer S.L., Sorger P.K. Measuring and modeling apoptosis in single cells. Cell 2011; 144(6): 926–39.
  5. Herr I., Debatin K.-M. Cellular stress response and apoptosis in cancer therapy. Blood 2001; 98(9): 2603–14.
  6. Chao D.T., Korsmeyer S. J. BCL-2 family: regulators of cell death. Ann. Rev. Immunol. 1998; 16: 395–419.
  7. Green D.R. Apoptotic pathways: The roads to ruin. Cell 1998; 94: 695–8.
  8. Chauncey T.R. Drug resistance mechanisms in acute leukemia. Curr. Opin. Oncol. 2001; 13(1): 21–6.
  9. Del Poeta G., Bruno A., Del M.I. Principe et al. Deregulation of the mitochondrial apoptotic machinery and development of molecular targeted drugs in acute myeloid leukemia. Curr. Cancer Drug Targets 2008; 8(3): 202–7.
  10. Testa U., Riccioni R. Deregulation of apoptosis in acute myeloid leukemia. Haematologica 2007; 92(1): 81–94.
  11. Banker D.E., Groudine M., Norwood T., Appelbaum F.R. Measurement of spontaneous and therapeutic agent-induced apoptosis with BCL-2 protein expression in acute myeloid leukemia. Blood 1997; 89(1): 243–55.
  12. Stijn A., Pol M.A., Kok A. et al. Differences between the CD34+ and CD34– blast compartments in apoptosis resistance in acute myeloid leukemia. Haematologica 2003; 88(5): 497–508.
  13. Fulda S., Los M., Friesen C., Debatin K.M. Chemosensitivity of solid tumor cells in vitro is related to activation of the CD95 system. Int. J. Cancer 1998; 76(1): 105–14.
  14. Fulda S., Sieverts H., Friesen C. et al. The CD95 (APO-1/Fas) system mediates drug-induced apoptosis in neuroblastoma cells. Cancer Res. 1997; 57(17): 3823–9.
  15. Барышников А.Ю., Полосухина Е.Р., Шишкин Ю.В. и др. Новый прогностический маркер острого лимфобластного лейкоза — антиген CD95 (FAS/APO-1). Гематол. и трансфузиол. 1998; 2: 8–11. [Baryshnikov A.Yu., Polosukhina Ye.R., Shishkin Yu.V. i dr. Novyy prognosticheskiy marker ostrogo limfoblastnogo leykoza — antigen CD95 (FAS/APO-1) (New prognostic marker of acute lymphoblastic leukemia — CD95 antigen (FAS/ APO-1). In: Hematol. & transfusiol.) Gematol. i transfuziol. 1998; 2: 8–11.]
  16. Molica S., Mannella A., Dattilo A. et al. Differential expression of BCL-2 oncoprotein and Fas antigen on normal peripheral blood and leukemic bone marrow cells. A flow cytometric analysis. Haematologica 1996; 81(4): 302–9.
  17. Greenblatt M.S., Bennett W.P., Hollstein M., Harris C.C. Mutations in the p53 tumor suppressor gene: clues to cancer etiology and molecular pathogenesis. Cancer Res. 1994; 54(18): 4855–78.
  18. Ko L.J., Prives C. p53: puzzle and paradigm. Genes & Dev. 1996; 10(9): 1054–72.
  19. Levine A.J. p53, the cellular gatekeeper for growth and division. Cell 1997; 88(3): 323–31.
  20. Resnick M.A., Tomso D., Inga A. et al. Functional diversity in the gene network controlled by the master regulator p53 in humans. Cell. Cycle 2005; 4(8): 1026–9.
  21. Prokocimer M., Rotter V. Structure and function of p53 in normal cells and their aberrations in cancer cells: projection on the hematologic cell lineages. Blood 1994; 84(8): 2391–411.
  22. Wada M., Bartram C.R., Nakamura H. et al. Analysis of p53 mutations in a large series of lymphoid hematologic malignancies of childhood. Blood 1993; 82(10): 3163–9.
  23. Cavalcanti G.B., Scheiner M.A., Simoes Magluta E.P. et al. p53 flow cytometry evaluation in leukemias: correlation to factors affecting clinical outcome. Cytometry B. Clin. Cytom. 2010; 78(4): 253–9.
  24. Beyazit Y., Aksu S., Haznedaroglu I.C. et al. Overexpression of the local bone marrow renin-angiotensin system in acute myeloid leukemia. J. Natl. Med. Assoc. 2007; 99: 57–63.
  25. Goker H., Haznedaroglu I.C., Beyazit Y. et al. Local umbilical cord blood renin-angiotensin system. Ann. Hematol. 2005; 84: 277–81.
  26. Jokubaitis V.J., Sinka L., Driessen R. et al. Angiotensin-converting enzyme (CD143) marks hematopoietic stem cells in human embryonic, fetal, and adult hematopoietic tissues. Blood 2008; 111: 4055–63.
  27. Li J., Volkov L., Comte L. et al. Production and consumption of the tetrapeptide AcSDKP, a negative regulator of hematopoietic stem cells, by hematopoietic microenvironmental cells. Exp. Hematol. 1997; 25: 140–6.
  28. Гальцева И.В., Пашин Л.Е., Савченко В.Г. Лейкемические дендритные клетки. Тер. арх. 2008; 80(7): 84–8. [Galtseva I.V., Pashin L.Ye., Savchenko V.G. Leykemicheskiye dendritnyye kletki (Leukemic dendritic cells. In: Ther. archive). Ter. arkh. 2008; 80(7): 84–8.]
  29. Danilov S.M., Sadovnikova E., Scharenborg N. et al. Angiotensinconverting enzyme (CD 143) is abundantly expressed by dendritic cells and discriminates human monocyte-derived dendritic cells from acute myeloid leukemia-derived dendritic cells. Exp. Hematol. 2003; 31(12): 1301–9.
  30. Del Poeta G., Venditti A., Del Principe M.I. et al. Amount of spontaneous apoptosis detected by Bax/Bcl-2 ratio predicts outcome in acute myeloid leukemia (AML). Blood 2003; 101(6): 2125–31.
  31. El-Manallawy H.A., El-Shkankiry N.H., El-Guindy S. et al. The Expression of Bcl-2 and Bax Proteins and Their Clinical Relevance in ALL and CLL Patients. J. Egypt. Nat. Cancer Inst. 2001; 13(1): 35–42.
  32. Hogarth L.A., Hall A.G. Increased BAX expression is associated with an increased risk of relapse in childhood acute lymphocytic leukemia. Blood 1999; 93(8): 2671–8.
  33. Aksu S., Beyazit Y., Haznedaroglu I.C. et al. Enhanced expression of the local haematopoietic bone marrow renin-angiotensin system in polycythemia rubra vera. J. Int. Med. Res. 2005; 33(6): 661–7.
  34. Aksu S., Beyazit Y., Haznedaroqlu I.C. et al. Over-expression of angiotensin-converting enzyme (CD 143) on leukemic blasts as a clue for the activated local bone marrow RAS in AML. Leuk. Lymphoma 2006; 47(5): 891–6.
  35. Барышников А.Ю., Шишкин Ю.В. Иммунологические проблемы апоптоза. М., 2002.  [Baryshnikov A.Yu., Shishkin Yu.V. Immunologicheskiye problemy apoptoza (Immunological problems of apoptosis). , 2002.]
  36. Miyawaki T., Uehara T., Nibu R. et al. Differential expression of apoptosisrelated Fas antigen on lymphocyte subpopulations in human peripheral blood. J. Immunol. 1992; 149(11): 3753–8.
  37. Lechner H., Amort M., Steger M.M. et al. Regulation of CD95 (APO-1) expression and the induction of apoptosis in human T cells: changes in old age. Int. Arch. Allergy Immunol. 1996; 110(3): 238–43.
  38. Kotani T., Aratake Y., Kondo S. et al. Expression of functional Fas antigen on adult T-cell leukemia. Leuk. Res. 1994; 18(4): 305–10.