Disulfiram targeting lymphoid malignant cell lines via ROS-JNK activation as well as Nrf2 and NF-kB pathway inhibition

Journal of Translational Medicine

Bing Xu¹  Weiguang Wang²  Peng Li³  Shiyun Wang¹  Rongwei Li¹  Yanyan Zhang¹  Yao Yao⁴  Pengcheng Shi¹  Huijuan Dong¹  Feili Chen¹  Jie Zha¹ 

[1] Department of Hematology, Nanfang Hospital, Southern Medical University, Guangzhou 510515, China;Research Institute in Healthcare Science, Faculty of Science and Engineering, University of Wolverhampton, Wolverhampton, UK;Drug Discovery Pipeline, Guangzhou Institutes of Biomedicine and Health, Chinese Academy of Sciences, Guangzhou 510530, China;Key Laboratory of Regenerative Biology, South China Institute for Stem Cell Biology and Regenerative Medicine, Guangzhou Institutes of Biomedicine and Health, Chinese Academy of Sciences, Guangzhou 510530, China

关键词: Apoptosis;    JNK;    NF-κB;    Nrf2;    Disulfiram;   

Others  :  810516
DOI  :  10.1186/1479-5876-12-163

 received in 2013-11-19, accepted in 2014-06-03,  发布年份 2014

【 摘 要 】

Background

Disulfiram (DS), an anti-alcoholism drug, demonstrates strong antitumor activity in a copper (Cu)-dependent manner. This study investigates the cytotoxicity of DS/Cu complex in lymphoid malignant cell lines in vitro and in vivo.

Method

Raji cells were subjected to different treatments and thereafter MTT assay, flow cytometry were used to determine IC₅₀ and apoptotic status. We also tested the cytotoxicity of DS/Cu in acute lymphoblastic leukemia cell line Molt4 in vitro. In vivo experiments were also performed to demonstrate the anticancer efficacy of DS/Cu in Raji cells xenografted nude mice.

Results

In combination with a low concentration (1 μM) of Cu²⁺, DS induced cytotoxicity in Raji cells with an IC₅₀ of 0.085 ± 0.015 μM and in Molt4 cells with an IC₅₀ of 0.435 ± 0.109 μM. The results of our animal experiments also showed that the mean tumor volume in DS/Cu-treated mice was significantly smaller than that in DS or control group, indicating that DS/Cu inhibits the proliferation of Raji cells in vivo. DS/Cu also induced apoptosis in 2 lymphoid malignant cell lines. After exposure to DS (3.3 μM)/Cu (1 μM) for 24 hours, apoptosis was detected in 81.03 ± 7.91% of Raji cells. DS/Cu induced significant apoptosis in a concentration-dependent manner with the highest apoptotic proportion (DS/Cu: 89.867 ± 4.69%) at a concentration of 2 μM in Molt4 cells. After 24 h exposure, DS/Cu inhibits Nrf2 expression. Flow cytometric analysis shows that DS/Cu induced ROS generation. DS/Cu induced phosphorylation of JNK and inhibits p65 expression as well as Nrf2 expression both in vitro and in vivo. N-acetyl-L-cysteine (NAC), an antioxidant, can partially attenuate DS/Cu complex-induced apoptosis and block JNK activation in vitro. In addition, NAC is able to restore Nrf2 nuclear translocation and p65 expression.

Conclusion

Our study manifests that DS/Cu complex targets lymphoid malignant cells in vitro and in vivo. Generation of ROS might be one of core steps in DS/Cu induced apoptosis. Moreover, ROS-related activation of JNK pathway and inhibition of NF-κB and Nrf2 may also contribute to the DS/Cu induced apoptosis.

【 授权许可】

   
2014 Zha et al.; licensee BioMed Central Ltd.

【 图 表 】

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