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Sensitivity and Resistance to Therapy

Characterization of the ubiquitin–proteasome system in bortezomib-adapted cells

Leukemia volume 23, pages 1098–1105 (2009)Cite this article

Abstract

Resistance towards the proteasome inhibitor bortezomib is poorly understood. We adapted the HL-60, ARH-77 and AMO-1 cell lines (myeloid leukemia, plasmocytoid lymphoma, myeloma) to bortezomib exceeding therapeutic plasma levels, and compared characteristics of the ubiquitin–proteasome system, alternative proteases and the unfolded protein response (UPR) between adapted cells and parental lines. Adapted cells showed increased transcription rates, activities and polypeptide levels of the bortezomib-sensitive β5, but also of the β2 proteasome subunit and consistently retained elevated levels of active β1/β5-type proteasome subunits in the presence of therapeutic levels of bortezomib. Bortezomib-adapted HL-60 cells showed increased expression and proteasome association of the 11S proteasome activator, and did not accumulate poly-ubiquitinated protein, activate the UPR or UPR-mediated apoptosis in response to bortezomib. The rate of protein biosynthesis was reduced, and the transcription of chaperone genes downmodulated. We did not observe major changes in the activities of TPPII, cathepsins or deubiquitinating proteases. We conclude that different types of bortezomib-adapted cell lines, including myeloma, show similar patterns of changes in the proteasomal machinery which result in residual proteasome activity in the presence of bortezomib and a quantitative balance between protein biosynthesis and destruction.

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Acknowledgements

We thank the Proteom-Centrum at the Interfaculty Institute for Cell Biology, University of Tübingen (Professor A Nordheim) for use of the fluorescence scanner and Carsten Henneges for help with data analysis. Research was supported by grants from Deutsche Forschungsgemeinschaft (SFB685), the Deutsche Krebshilfe and the Swiss National Science Foundation (to CD) as well as the Clinical Trials Unit at the Cantonal Hospital St Gallen (CTU-KSSG).

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Authors and Affiliations

  1. Department of Medicine II, University of Tübingen, Tübingen, Germany

    T Rückrich, M Kraus, J Gogel & C Driessen

  2. ZfKM GmbH, Heilbronn, Germany

    A Beck

  3. Division of Cell Biology II, The Netherlands Cancer Institute, Amsterdam, The Netherlands

    H Ovaa

  4. Institute of Chemistry, Bio-organic Synthesis, Leiden University, Leiden, The Netherlands

    M Verdoes & H S Overkleeft

  5. Interfaculty Institute of Biochemistry, University of Tübingen, Tübingen, Germany

    H Kalbacher

  6. Department of Oncology and Hematology, Cantonal Hospital St Gallen, St Gallen, Switzerland

    C Driessen

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  1. T Rückrich
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Correspondence to C Driessen.

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Supplementary Information accompanies the paper on the Leukemia website (http://www.nature.com/leu)

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Rückrich, T., Kraus, M., Gogel, J. et al. Characterization of the ubiquitin–proteasome system in bortezomib-adapted cells. Leukemia 23, 1098–1105 (2009). https://doi.org/10.1038/leu.2009.8

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