Blood Res.  2015 Mar;50(1):33-39. 10.5045/br.2015.50.1.33.

Anti-leukemic properties of deferasirox via apoptosis in murine leukemia cell lines

• Jeon SR
• Lee JW
• Jang PS
• Chung NG
• Cho B
• Jeong DC

• KMID: 1787910
• DOI: http://doi.org/10.5045/br.2015.50.1.33

Abstract

BACKGROUND
Although deferasirox (DFX) is reported to have anti-tumor effects, its anti-leukemic activity remains unclear. We evaluated the effect of DFX treatment on two murine lymphoid leukemia cell lines, and clarified the mechanisms underlying its potential anti-leukemic activity.
METHODS
L1210 and A20 murine lymphoid leukemia cell lines were treated with DFX. Cell viability and apoptosis were evaluated by the 3-(4,5-dimethylthaizol-2-yl)-5-(3-carboxymethylphenyl)-2-(4-sulfophenyl)-2H-tetrazolium (MTS) assay and fluorescence-activated cell sorting (FACS) analysis, respectively. Immunoblotting was performed to detect the expression of key apoptotic proteins.
RESULTS
In dose- and time-dependent manner, DFX decreased viability and increased apoptosis of murine leukemic cells. Fas expression was significantly higher in A20 cells than in L1210 cells at all DFX concentrations tested. Although both cell lines exhibited high caspase 3 and caspase 9 expression, a critical component of the intrinsic mitochondrial apoptotic pathway, expression was greater in L1210 cells. In contrast, caspase 8, a key factor in the extrinsic apoptotic pathway, showed greater expression in A20 cells. Cytochrome c expression was significantly higher in L1210 cells. In both cell lines, co-treatment with ferric chloride and DFX diminished the expression of these intracellular proteins, as compared to DFX treatment alone.
CONCLUSION
Treatment with DFX increased caspase-dependent apoptosis in two murine lymphoid leukemia cell lines, with differing apoptotic mechanisms in each cell line.

Keyword

Lymphoid leukemia; Deferasirox; Apoptosis; Caspase

MeSH Terms

Apoptosis*
Caspase 3
Caspase 8
Caspase 9
Cell Line*
Cell Survival
Cytochromes c
Flow Cytometry
Immunoblotting
Leukemia*
Leukemia, Lymphoid
Caspase 3
Caspase 8
Caspase 9
Cytochromes c

Figure

• 

  Fig. 1 Viability of L1210 (A, B) and A20 cells (C, D) after DFX treatment with or without FeCl3, according to treatment time (24 vs. 48 hr) and DFX concentration (3.125-75 µM). Experiments were performed in triplicate. All data are presented as the mean±standard error (a)P<0.05, b)P<0.01).

• 

  Fig. 2 Measurement of apoptosis in L1210 (A, B) and A20 cells (C, D) after DFX treatment with or without FeCl3, according to treatment time (24 vs. 48 hr) and DFX concentration (12.5-50 µM). Experiments were performed in triplicate. All data are presented as the mean±standard error (a)P<0.05, b)P<0.01).

• 

  Fig. 3 Measurement of CD95 (Fas) expression in L1210 and A20 cells after 24 h DFX treatment with or without FeCl3, according to DFX concentration (12.5-50 µM). Experiments were performed in triplicate. All data are presented as the mean±standard error (a)P<0.05, b)P<0.01).

• 

  Fig. 4 Western blot measurement of caspase 3, caspase 9, caspase 8, PARP, and BAX expression in L1210 (A) and A20 cells (B) after treatment with 50 µM DFX with or without FeCl3, according to treatment time (6-48 hr).

• 

  Fig. 5 Western blot measurement of cytochrome c expression in L1210 (A) and A20 cells (B) after treatment with 50 µM DFX with or without FeCl3, according to treatment time (6-48 hr).

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