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

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 a...

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Autores principales: Zha, Jie, Chen, Feili, Dong, Huijuan, Shi, Pengcheng, Yao, Yao, Zhang, Yanyan, Li, Rongwei, Wang, Shiyun, Li, Peng, Wang, Weiguang, Xu, Bing
Formato: Online Artículo Texto
Lenguaje:English
Publicado: BioMed Central 2014
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4075939/
https://www.ncbi.nlm.nih.gov/pubmed/24915933
http://dx.doi.org/10.1186/1479-5876-12-163
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author Zha, Jie
Chen, Feili
Dong, Huijuan
Shi, Pengcheng
Yao, Yao
Zhang, Yanyan
Li, Rongwei
Wang, Shiyun
Li, Peng
Wang, Weiguang
Xu, Bing
author_facet Zha, Jie
Chen, Feili
Dong, Huijuan
Shi, Pengcheng
Yao, Yao
Zhang, Yanyan
Li, Rongwei
Wang, Shiyun
Li, Peng
Wang, Weiguang
Xu, Bing
author_sort Zha, Jie
collection PubMed
description 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 IC(50) 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 Cu(2+), DS induced cytotoxicity in Raji cells with an IC(50) of 0.085 ± 0.015 μM and in Molt4 cells with an IC(50) 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.
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spelling pubmed-40759392014-07-01 Disulfiram targeting lymphoid malignant cell lines via ROS-JNK activation as well as Nrf2 and NF-kB pathway inhibition Zha, Jie Chen, Feili Dong, Huijuan Shi, Pengcheng Yao, Yao Zhang, Yanyan Li, Rongwei Wang, Shiyun Li, Peng Wang, Weiguang Xu, Bing J Transl Med Research 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 IC(50) 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 Cu(2+), DS induced cytotoxicity in Raji cells with an IC(50) of 0.085 ± 0.015 μM and in Molt4 cells with an IC(50) 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. BioMed Central 2014-06-11 /pmc/articles/PMC4075939/ /pubmed/24915933 http://dx.doi.org/10.1186/1479-5876-12-163 Text en Copyright © 2014 Zha et al.; licensee BioMed Central Ltd. http://creativecommons.org/licenses/by/2.0 This is an Open Access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/2.0), which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly credited. The Creative Commons Public Domain Dedication waiver (http://creativecommons.org/publicdomain/zero/1.0/) applies to the data made available in this article, unless otherwise stated.
spellingShingle Research
Zha, Jie
Chen, Feili
Dong, Huijuan
Shi, Pengcheng
Yao, Yao
Zhang, Yanyan
Li, Rongwei
Wang, Shiyun
Li, Peng
Wang, Weiguang
Xu, Bing
Disulfiram targeting lymphoid malignant cell lines via ROS-JNK activation as well as Nrf2 and NF-kB pathway inhibition
title Disulfiram targeting lymphoid malignant cell lines via ROS-JNK activation as well as Nrf2 and NF-kB pathway inhibition
title_full Disulfiram targeting lymphoid malignant cell lines via ROS-JNK activation as well as Nrf2 and NF-kB pathway inhibition
title_fullStr Disulfiram targeting lymphoid malignant cell lines via ROS-JNK activation as well as Nrf2 and NF-kB pathway inhibition
title_full_unstemmed Disulfiram targeting lymphoid malignant cell lines via ROS-JNK activation as well as Nrf2 and NF-kB pathway inhibition
title_short Disulfiram targeting lymphoid malignant cell lines via ROS-JNK activation as well as Nrf2 and NF-kB pathway inhibition
title_sort disulfiram targeting lymphoid malignant cell lines via ros-jnk activation as well as nrf2 and nf-kb pathway inhibition
topic Research
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4075939/
https://www.ncbi.nlm.nih.gov/pubmed/24915933
http://dx.doi.org/10.1186/1479-5876-12-163
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