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 1

NF-𝜅B, c-Jun, p53, and caspase-3 signalling molecules and oxidative stress are involved in TPEN-induced apoptosis in Jurkat cells. Jurkat cells (1 × 10⁶ cells/mL) were exposed to TPEN (3 μM) in absence or presence of PDTC (10 nM), pifithrin-𝛼 (PFT, 50 nM), SP600125 (1 μM), and NSCI (10 μM) signaling inhibitors, and ZnSO₄ (10 μM) and NAC (1 mM) at 37°C for 24 h. Cells were then evaluated for apoptotic nuclei morphology, plasma membrane integrity, and mitochondrial membrane potential, as described in Material and Methods section. The percentage of positive AO/EB/Hoechst staining, AV⁺/7-AAD^−/+ and PI^−/+/DiOC₆(3)⁻ of Jurkat cells treated with or without TPEN expressed as a mean percentage ± S.D. from three independent experiments. ^∗∗𝑃<0.05.

Treatment/assay	AO/EB/Hoechst (%)	AV^+/7-AAD^−/+ (%)	PI^−/+/DiOC6(3)^− (%)
Untreated	<1 ± 0	4±1	3±1
TPEN (3 𝜇M)	100	82±2	80±1
PDTC (10 nM)	<1 ± 0	2±1	3±2
PDTC (10 nM) + TPEN (3 𝜇M)	5±1^∗∗	18±1^∗∗	24±2^∗∗
SP600125 (1 μM)	<1 ± 1	3±1	2±1
SP600125 (1 μM) + TPEN (3 μM)	3±1^∗∗	10±1^∗∗	39±2^∗∗
PFT (50 nM)	<1 ± 0	2±1	3±1
PFT (50 nM) + TPEN (3 μM)	6±1^∗∗	28±1^∗∗	20±1^∗∗
NSCI (10 μM)	<1 ± 0	3±1	2±2
NSCI (10 μM) + TPEN (3 μM)	8±1^∗∗	33±1^∗∗	35±1^∗∗
ZnSO4 (10 μM)	<1 ± 0	1±1	1±1
ZnSO4 (10 μM) + TPEN (3 μM)	3±2^∗∗	6±1^∗∗	6±1^∗∗
Zn (10 μM)/TPEN (3 μM) complex	6±2^∗∗	5±1^∗∗	3±1^∗∗
NAC (1 mM)	<1 ± 0	3±2	2±2
NAC (1 mM) + TPEN (3 μM)	2±1^∗∗	10±3^∗∗	15±2^∗∗