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BCNU-sequestration by metallothioneins may contribute to resistance in a medulloblastoma cell line

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Abstract

Purpose

Resistance of neoplastic cells to the alkylating drug BCNU [1,3-bis(2-chloroethyl)-1-nitrosourea] has been correlated with expression of O 6-methylguanine-DNA methyltransferase, which repairs the O 6-chloroethylguanine produced by the drug. Other possible mechanisms of resistance include raised levels of glutathione or increased repair of the DNA interstrand cross-links formed by BCNU. Transcriptional profiling revealed the upregulation of several metallothionein (MT) genes in a BCNU-resistant medulloblastoma cell line [D341 MED (OBR)] relative to its parental line. Previous studies have shown that MTs, through their reactive thiol groups can quench nitrogen mustard-derived alkylating drugs. In this report, we evaluate whether MTs can also quench BCNU.

Methods

To demonstrate the binding of BCNU to MT, we used an assay that measured the release of the MT-bound divalent cations (Zn2+, Cd2+) upon their displacement by the drug. We also measured the decomposition rates of BCNU at those reaction conditions.

Results

The rate of release of the cations was higher in pH 7.4 than at pH 7.0, which is likely a result of more rapid decomposition of BCNU (thus faster release of MT-binding intermediate) at pH 7.4 than at pH 7.0.

Conclusion

We demonstrate that resistance to BCNU may be a result of elevated levels of MTs which act by sequestering the drug’s decomposition product(s).

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Acknowledgments

This work was supported by NIH grants NS-30245 and NS-20023 and a grant from the Pediatric Brain Tumor Foundation.

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Author notes

  1. Manny D. Bacolod

    Present address: Department of Microbiology and Immunology, Cornell University Weill Medical College, New York, NY, 10021, USA

Authors and Affiliations

  1. Department of Surgery, Duke University Medical Center, Durham, NC, 27710, USA

    Manny D. Bacolod, Stewart P. Johnson, Nancy S. Bullock & Henry S. Friedman

  2. Department of Medicine, Duke University Medical Center, Durham, NC, 27710, USA

    Randy Fehdrau & Michael Colvin

  3. Department of Pathology, Duke University Medical Center, Durham, NC, 27710, USA

    Darell D. Bigner

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  1. Manny D. Bacolod
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Correspondence to Manny D. Bacolod.

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Bacolod, M.D., Fehdrau, R., Johnson, S.P. et al. BCNU-sequestration by metallothioneins may contribute to resistance in a medulloblastoma cell line. Cancer Chemother Pharmacol 63, 753–758 (2009). https://doi.org/10.1007/s00280-008-0792-9

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  • DOI: https://doi.org/10.1007/s00280-008-0792-9

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