Publication Date:
2013-11-15
Description:
Introduction Chronic lymphocytic leukemia (CLL) shows a conspicuous inhomogeniety in the clinical course among patient subgroups with pronounced genetic aberrations. Some patients have a rapidly fatal disease despite therapy, which is accompanied by the incidence of deletion of chromosome 17p and mutation of TP53. Searching for new therapeutic options we investigate the cytotoxic effects and mechanistically opportunities of NO-ASA on CLL cells. Methods Primary cells were isolated from peripheral blood of CLL patients or healthy individuals. Primary CLL cells with or without TP53 mutation, as well as PBMCs and B-cells of healthy volunteers were tested in a luminometric assay (ATP content) after 24 h NO-ASA incubation. Caspase-dependency was determined in a luminometric assay after 6 h (n=5) as well as in immunoblot-analysis (Caspase 3, PARP; n=3) after 24 h. Influence on Btk- and NF-κB pathways was investigated after 3 h via immunoblot-analysis. Additional, expressions of NF-κB target genes (BCl-2, MCl-1 and CFLAR) were investigated via qRT-PCR after 4 h. Results NO-ASA effectively reduced ATP content in CLL cells from a mixed patient population (LD50 4.34 µM). Primary CLL cells from TP53 mutated patients (n=5) showed a slightly increased LD50 of 25.56 µM, which was still significantly lower than for healthy PBMCs (LD50 63.72 µM, n=5). In addition a concentration-dependent activation of the caspase cascade could be shown compared to untreated control (p
Print ISSN:
0006-4971
Electronic ISSN:
1528-0020
Topics:
Biology
,
Medicine
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