Abstract
Bortezomib is a proteasome inhibitor for the treatment of relapsed/refractory multiple myeloma (MM). Mechanisms of resistance to Bortezomib are undefined. Myeloid cell leukemia-1 (Mcl-1) is an antiapoptotic protein, which protects tumor cells against spontaneous and chemotherapy-induced apoptosis. In MM, specific downregulation of Mcl-1 induces apoptosis. Here, we examined the role of Mcl-1 in Bortezomib- and doxorubicin-induced apoptosis. We demonstrate that Bortezomib, but not doxorubicin, triggers caspase-dependent generation of a 28 kDa Mcl-1-fragment, in several MM cell lines, including MM.1S cells. Conversely, transient transfection of MM.1S cells with a previously reported 28 kDa Mcl-1128–350 fragment, but not with the Mcl-11–127 fragment, induces apoptosis. Therefore, both downregulation of full-length antiapoptotic Mcl-1, as well as Bortezomib-induced generation of Mcl-1128–350 cleaved protein, contribute to MM cell apoptosis. To verify further these findings, we next compared effects triggered by Bortezomib, doxorubicin and melphalan in Mcl-1wt/wt and Mcl-1Δ/null murine embryonic fibroblasts (MEFs). Our results show that Bortezomib, but not doxorubicin or melphalan, triggers Mcl-1 cleavage in Mcl-1wt/wt, but not Mcl-1Δ/null MEFs and induces sub-G1 phase cells; caspase-3 and -9, and PARP cleavage as well as morphological signs of apoptosis. Taken together, these results support an important role of Mcl-1 and a Mcl-1 fragment in Bortezomib-induced cell death in general, and in MM in particular. To prevent relapse of MM in patients treated with Bortezomib, we therefore recommend the combination of Bortezomib with agents that induce MM cell death independent of Mcl-1.
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Acknowledgements
We are grateful for reagents provided by G Packham (University of Southampton School of Medicine, Southampton General Hospital, Southampton, UK). K Podar is the recipient of the Dunkin Donut's Rising Star Award 2007. This work was supported by a Multiple Myeloma Foundation (MMRF) Senior Research Grant Award (KP), National Institute of Health Grants RO CA 50947, PO-1 CA 78378, and P50 CA 10070; and the Doris Duke Distinguished Clinical Research Scientist Award (KCA).
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Podar, K., Gouill, S., Zhang, J. et al. A pivotal role for Mcl-1 in Bortezomib-induced apoptosis. Oncogene 27, 721–731 (2008). https://doi.org/10.1038/sj.onc.1210679
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DOI: https://doi.org/10.1038/sj.onc.1210679
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