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Up-regulation of γ-glutamylcysteine synthetase activity in melphalan-resistant human multiple myeloma cells expressing increased glutathione levels

  • Original Articles
  • Glutathione, Melphalan, γ-Glutamylcysteine Synthetase
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Abstract

Levels of intracellular glutathione (GSH) and the GSH-related enzymes γ-glutamylcysteine synthetase (γ-GCS) and γ-glutamyltranspeptidase (γ-GT) were measured in the melphalan-resistant human multiple myeloma cell line 8226/LR-5 and were compared to those measured in the drug-sensitive 8226/S and doxorubicin-resistant 8226/Dox40 cell lines. Both GSH and γ-GCS activity, the rate-limiting step in the de novo synthesis of GSH, were elevated by a factor of approximately 2 in the melphalanresistant 8226/LR-5 cells relative to the other two lines. γ-GT activity was not elevated significantly in the /LR-5 cells. Northern analysis with a probe specific for the large subunit of human liver γ-GCS identified two bands (3.2 and 4.0 kb), both of which were increased by a factor of 2–3 in the 8226/LR-5 line. Levels of γ-GCS mRNA expression were comparable in the /S and /Dox40 cell lines. Levels of γ-GT mRNA were similar in the /S and /LR-5 lines but were reduced in the /Dox40 cells. These data suggest that the increased GSH levels associated with resistance to melphalan in the 8226/LR-5 myeloma cells is attributable to up-regulation of γ-GCS. This observation is consistent with recent demonstrations of up-regulation of γ-GCS in melphalan-resistant prostate carcinoma cells and cisplatinum-resistant ovarian carcinoma cells, suggesting that increased expression of γ-GCS may be an important mediator of GSH-associated resistance mechanisms.

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Authors and Affiliations

  1. Department of Human Oncology, University of Wisconsin-Madison, 600 Highland Avenue, K4/338, 53792, Madison, WI, USA

    R. Timothy Mulcahy, Howard H. Bailey & Jerry J. Gipp

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  1. R. Timothy Mulcahy
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  2. Howard H. Bailey
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  3. Jerry J. Gipp
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Mulcahy, R.T., Bailey, H.H. & Gipp, J.J. Up-regulation of γ-glutamylcysteine synthetase activity in melphalan-resistant human multiple myeloma cells expressing increased glutathione levels. Cancer Chemother. Pharmacol. 34, 67–71 (1994). https://doi.org/10.1007/BF00686114

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  • DOI: https://doi.org/10.1007/BF00686114

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