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Effects of Magnetic Nanoparticle of Fe
3
O
4
and
5Bromotetrandrin on apoptosis induced by
Daunorubicin in K562/A02 Leukemic cells
Shen Mingfang
1
, Chen Baoan
1*
, Cheng Jian
1
, Gao Feng
1
, Xu Wenlin
2
, Ding Jiahua
1
,
Gao Chong
1
, Sun Xinchen
3
, Li Guohong
1
, Chen Wenji
1
, Liu Lijie
4
, Li Xiaomao
5
,
and Wang Xuemei
6
1
Department of Hematology, Zhongda Hospital, Clinical medical school, Southeast University,
Nanjing (210009)
2
Department of Hematology, People’s Hospital, Jiangsu University ,Zhenjiang, 212002, China
3
Department of Oncology, Zhongda Hospital, Clinical medical school, Southeast University,
Nanjing (210009)
4
Institution of Physiology, Southeast University, Nanjing, 210009, China
5
Department of Physics, University of Saarland, D-66041 Saarbruechen, Germany
6
National Key Lab of Bioelectronics(Chien-Shiung Wu Laboratory), Southeast University,
Nanjing (210009)
E-mail: cba8888@hotmail.com
Abstract
The aim of this study was to investigate the potential benefit of combination therapy with Magnetic
Nanoparticle of Fe
3
O
4
and 5Bromotetrandrine on chronic leukemia. We detected the apoptosis by flow
cytometry(FCM) and Wright staining. The expression of BAX and BCL-2 was measured by Western
blot.The results showed that combination of DNR with either MNP(Fe
3
O
4
) or 5-BrTet exerted a potent
cytotoxic effect on K562/A02 cells, while MNP(Fe
3
O
4
) and 5-BrTet cotreatment could synergistically
enhance DNR-induced apoptosis. After treated with this composite, K562/A02 cells appeared typical
apoptotic morphological features; the expression level of BCL-2 decreased and BAX increased
markedly.It is concluded that MNP(Fe
3
O
4
) or 5-BrTet with DNR induced apoptosis in K562/ A02 cells,
and when used together,they had distinct synergism .The down-regulation of BCL-2 and the
up-regulation of BAX may play important roles.
Keywords:K562/A02 leukemic cells; multidrug resistance; Magnetic Nanoparticle of
Fe
3
O
4
;5Bromotetrandrine; apoptosis; BCL-2;BAX
1. Introduction
Most cytotomic agents exert their anticancer effects by inducing apoptotic- programmed cell
death
[ 1 ]
, the resistance of leukemic cells to chemotherapy-induced apoptosis remains the most
significant problem in the development of drug resistance and treatment failure in acute leukemia
[ 2 ]
.
To reverse the resistance mechanism and reduce side effects during chemotherapy, a promising
approach is to combine a conventional chemotherapy with new strategies inducing leukemia cells
apoptosis. In our previous study, it was found that 5Bromotetrandrine (5-BrTet) could effectively
reverse P-glycoprotein (P-gp)-mediated multidrug resistance
[ 3 ]
,and MNP(Fe
3
O
4
) loaded with
chemotherapeutic agent and chemosensitizer had synergetic effect on reveral in multidrug resistance
[ 4 ]
.
The present studies were undertaken to evaluate the ability of MNP(Fe
3
O
4
) and 5-BrTet to sensitize
MDR cells to anticancer agents in vitro, to determine their combination effects on apoptosis induced by
combining DNR in leukemic cells.
2. Materials and methods
2.1 Main reagents
Adriamycin (ADM)( Hisun Phamaceutical Co., Zhejiang, China) and Daunorubicin (DNR)(Main
Luck Phamaceuticals Inc., Shenzheng, China) stock solution 2mg/ml were prepared with 0.01mol/l
phosphate buffer saline (PBS)(PH 7.4). 5Bromotetrandrine (5-BrTet) (Kanghong, Chengdou, China)