Abstract
Introduction
Imatinib, a small-molecule inhibitor of the Bcr-Abl kinase, is a successful drug for treating chronic myeloid leukemia (CML). Bcr-Abl kinase stimulates the production of H2O2, which in turn activates Abl kinase. We therefore evaluated whether N-acetyl cysteine (NAC), a ROS scavenger improves imatinib efficacy.
Materials and methods
Effects of imatinib and NAC either alone or in combination were assessed on Bcr-Abl+ cells to measure apoptosis. Role of nitric oxide (NO) in NAC-induced enhanced cytotoxicity was assessed using pharmacological inhibitors and siRNAs of nitric oxide synthase isoforms. We report that imatinib-induced apoptosis of imatinib-resistant and imatinib-sensitive Bcr-Abl+ CML cell lines and primary cells from CML patients is significantly enhanced by co-treatment with NAC compared to imatinib treatment alone. In contrast, another ROS scavenger glutathione reversed imatinib-mediated killing. NAC-mediated enhanced killing correlated with cleavage of caspases, PARP and up-regulation and down regulation of pro- and anti-apoptotic family of proteins, respectively. Co-treatment with NAC leads to enhanced production of nitric oxide (NO) by endothelial nitric oxide synthase (eNOS). Involvement of eNOS dependent NO in NAC-mediated enhancement of imatinib-induced cell death was confirmed by nitric oxide synthase (NOS) specific pharmacological inhibitors and siRNAs. Indeed, NO donor sodium nitroprusside (SNP) also enhanced imatinib-mediated apoptosis of Bcr-Abl+ cells.
Conclusion
NAC enhances imatinib-induced apoptosis of Bcr-Abl+ cells by endothelial nitric oxide synthase-mediated production of nitric oxide.
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Abbreviations
- NAC:
-
N-acetyl cysteine
- CML:
-
Chronic myeloid leukemia
- NO:
-
Nitric oxide
- NOS:
-
Nitric oxide synthase
- ROS:
-
Reactive oxygen species
- SNP:
-
Sodium nitroprusside
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Acknowledgments
We thank Mr. Anirban Manna for preparation of the manuscript and Dr. Carlo Gambacorti Passerini for providing KU812 and KCL22 cell lines. The present study was supported by the Council of Scientific and Industrial Research (CSIR), Department of Biotechnology (DBT), New Delhi.
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Rakshit, S., Bagchi, J., Mandal, L. et al. N-acetyl cysteine enhances imatinib-induced apoptosis of Bcr-Abl+ cells by endothelial nitric oxide synthase-mediated production of nitric oxide. Apoptosis 14, 298–308 (2009). https://doi.org/10.1007/s10495-008-0305-7
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DOI: https://doi.org/10.1007/s10495-008-0305-7