Korean J Hematol.  2012 Sep;47(3):194-201. 10.5045/kjh.2012.47.3.194.

Effects of oral iron chelator deferasirox on human malignant lymphoma cells

• Choi JG
• Kim JL
• Park J
• Lee S
• Park SJ
• Kim JS
• Choi CW

• KMID: 2251953
• DOI: http://doi.org/10.5045/kjh.2012.47.3.194

Abstract

BACKGROUND
Iron is essential for cell proliferation and viability. It has been reported that iron depletion by a chelator inhibits proliferation of some cancer cells. Deferasirox is a new oral iron chelator, and a few reports have described its effects on lymphoma cells. The goal of this study was to determine the anticancer effects of deferasirox in malignant lymphoma cell lines.
METHODS
Three human malignant lymphoma cell lines (NCI H28:N78, Ramos, and Jiyoye) were treated with deferasirox at final concentrations of 20, 50, or 100 microM. Cell proliferation was evaluated by an MTT assay, and cell cycle and apoptosis were analyzed by flow cytometry. Western blot analysis was performed to determine the relative activity of various apoptotic pathways. The role of caspase in deferasirox-induced apoptosis was investigated using a luminescent assay.
RESULTS
The MTT assay showed that deferasirox had dose-dependent cytotoxic effects on all 3 cell lines. Cell cycle analysis showed that the sub-G1 portion increased in all 3 cell lines as the concentration of deferasirox increased. Early apoptosis was also confirmed in the treated cells by Annexin V and PI staining. Western blotting showed an increase in the cleavage of PARP, caspase 3/7, and caspase 9 in deferasirox-treated groups.
CONCLUSION
We demonstrated that deferasirox, a new oral iron-chelating agent, induced early apoptosis in human malignant lymphoma cells, and this apoptotic effect is dependent on the caspase-3/caspase-9 pathway.

Keyword

Deferasirox; Malignant lymphoma; Apoptosis

MeSH Terms

Annexin A5
Apoptosis
Benzoates
Blotting, Western
Caspase 9
Cell Cycle
Cell Line
Cell Proliferation
Flow Cytometry
Humans
Iron
Lymphoma
Triazoles
Annexin A5
Benzoates
Caspase 9
Iron
Triazoles

Figure

• 

  Fig. 1 MTT assay results show that deferasirox inhibited the proliferation of malignant lymphoma cell lines (A) NCI H28:N78 cells, (B) Jiyoye cells, and (C) Ramos cells. The results are presented as mean (SD) percentage of viability from triplicate cultures with repeated experiments (a)P<0.05, b)P<0.01).

• 

  Fig. 2 Flow cytometry analysis results show that deferasirox treatment increased the percentage of cells in sub-G1 phase in a dose-dependent manner. (A) NCI H28:N78 cells, (B) Jiyoye cells, and (C) Ramos cells. The results are presented as mean (SD) percentage of cells in sub-G1 from triplicate experiments (a)P<0.05, b)P<0.01).

• 

  Fig. 3 Deferasirox induces early apoptosis. Treatment with deferasirox caused dose-dependent apoptosis in 3 malignant lymphoma cell lines (A) NCI H28:N78 cells, (B) Jiyoye cells, and (C) Ramos cells. The experiments were repeated 3 times.

• 

  Fig. 4 Colorimetric assay results show that deferasirox treatment increased caspase-3/7 and caspase-9 activities. Caspase-3/7 and caspase-9 activities were measured after cells were treated with deferasirox for 24 h (A) NCI H28:N78 cells, (B) Jiyoye cells, and (C) Ramos cells (a)P<0.05, b)P<0.01).

• 

  Fig. 5 Western blot analysis of apoptosis-related modulators. Expression of Bax and P53 was upregulated, whereas the expression of Bcl-2 was downregulated after malignant lymphoma cell lines were treated with deferasirox.

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