Apoptosis assessment in acute myeloid leukemia and B-cell chronic lymphocytic leukemia cells after in vitro cytostatics treatment and its relationship to multidrug resistance

Česká verzia

Authors: K. Jankovičová ¹; J. Krejsek ¹; O. Kopecký ¹; J. Voglová ²; Z. Škrabková ³; J. Novosad ¹
Authors place of work: Ústav klinické imunologie a alergologie Lékařské fakulty UK a FN Hradec Králové, přednosta MUDr. Otakar Kopecký, CSc. ¹; Oddělení klinické hematologie FN Hradec Králové, přednosta prof. MUDr. Jaroslav Malý, CSc. ²; Ústav lékařské biofyziky Lékařské fakulty UK a FN Hradec Králové, přednosta prof. MUDr. Pravoslav Stránský, CSc. ³
Published in the journal: Vnitř Lék 2005; 51(2): 175-182
Category: Original Contributions

Summary

Apoptosis failure during chemotherapy of leukemia is one of the multidrug resistance mechanisms. The aim of this study was to introduce in vitro model for malignant cell clone sensitivity testing. In vitro cytostatic treatment of acute myeloid leukemia (AML) and B-cell chronic lymphocytic leukemia (B-CLL) cells and its relationship to P-gp, MRP, LRP, Bcl-2, and Bax proteins expression was also investigated. AML cells were treated with doxorubicin and B-CLL cells were treated with chlorambucil. Flow cytometry was used for all experiments evaluation. To detect the amount of apoptotic cells in the sample three methods were used: annexin test, TUNEL (Tdt-mediated dUTP nick end labeling) test, and caspase 8 detection. After in vitro cytostatic treatment statistically significant increase in the number of apoptotic cells in the culture in the case of AML cells but not in the case of B-CLL cells was observed. When analysing multidrug resistance markers some tendencies to correlate with in vitro apoptosis reaction of AML and B-CLL cells were found above all by P-gp and Bcl-2 markers. AML cells displayed negative correlation between P-gp function and number of apoptotic cells detected by TUNEL test. B-CLL cells demonstrated negative correlation between P-gp expression and number of apoptotic cells detected by annexin test and negative correlation between Bcl-2 and caspase 8 expression.

Key words:
acute myeloid leukemia – B-cell chronic lymphocytic leukemia – apoptosis – multidrug resistance – flow cytometry

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