Research Article

TPEN Induces Apoptosis Independently of Zinc Chelator Activity in a Model of Acute Lymphoblastic Leukemia and Ex Vivo Acute Leukemia Cells through Oxidative Stress and Mitochondria Caspase-3- and AIF-Dependent Pathways

Table 3

TPEN induces nuclei fragmentation, mitochondrial membrane depolarization, activation of p53, caspase-3, and AIF in isolated ALL cells. ALL cells were left untreated or exposed to TPEN (5  M) for 24 h. After this time, cells were evaluated for fluorescence apoptotic nuclei morphology by AO/EB/Hoechst staining, mitochondrial potential damage by PI/DiOC6(3) flow cytometry, and p53, caspase-3, and AIF detection by immunohistochemistry, as described in Material and Methods section. Percentage of positive staining markers of untreated or treated ALL cells with TPEN is expressed as a mean percentage ± S.D. of two replicas from one independent experiment.

TreatmentAssay
AO/EB/Hoechst (%)PI−/+/DiOC6(3) (%)p53 (%)Caspase-3 (%)AIF (%)

Untreated
5 ± 29 ± 39 ± 213 ± 216 ± 2
TPEN (5  M)
97 ± 179 ± 274 ± 390 ± 382 ± 3