Abstract
Bortezomib is an effective agent for treating multiple myeloma (MM). To investigate the underlying mechanisms associated with acquired resistance to this agent, we established two bortezomib-resistant MM cell lines, KMS-11/BTZ and OPM-2/BTZ, the 50% inhibitory concentration values of which were respectively 24.7- and 16.6-fold higher than their parental cell lines. No activation of caspase and BH3-only proteins such as Noxa was noted in bortezomib-resistant cells after exposure to the drug. The accumulation of polyubiquitinated proteins was reduced in bortezomib-resistant cells compared with the parental cells, associated with avoidance of catastrophic ER stress as assessed by downregulation of CHOP expression. These resistant MM cells have a unique point mutation, G322A, in the gene encoding the proteasome β5 subunit (PSMB5), likely resulting in conformational changes to the bortezomib-binding pocket of this subunit. KMS-11 parental cells transfected to express mutated PSMB5 also showed reduced bortezomib-induced apoptosis compared with those expressing wild-type PSMB5 or the parental cells. Expression of mutated PSMB5 was associated with the prevention of the accumulation of unfolded proteins. Thus, a fraction of MM cells may acquire bortezomib resistance by suppressing apoptotic signals through the inhibition of unfolded protein accumulation and subsequent excessive ER stress by a mutation of the PSMB5 gene.
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Acknowledgements
We thank Ms Chiori Fukuyama for her skillful technical assistance. This work was supported by Grants-in-Aid for Scientific Research on Priority Areas (No. 17016065 & 16062101 for RU) from the Ministry of Education, Culture, Science, Sports and Technology, Japan; and Grants-in-Aid for Cancer Research from the Ministry of Health, Labor and Welfare, Japan (No. 17S-1, 17-16 anf 21-8-5 for SI). This research was also funded in part by Kyowa Hakko Kirin Co., Ltd, Tokyo, Japan.
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TN, HM and YS are employees of Kyowa Hakko Kirin Co., Ltd., Japan. SI received research funding from Kyowa Hakko Kirin. SI declares honoraria from Janssen Pharmaceutical K.K., Dainippon Sumitomo Pharmaceutical Co., Ltd, Chugai Pharmaceutical Co., Ltd and Novartis Pharma K.K.
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Ri, M., Iida, S., Nakashima, T. et al. Bortezomib-resistant myeloma cell lines: a role for mutated PSMB5 in preventing the accumulation of unfolded proteins and fatal ER stress. Leukemia 24, 1506–1512 (2010). https://doi.org/10.1038/leu.2010.137
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DOI: https://doi.org/10.1038/leu.2010.137
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