Adaptive response to GSH depletion and resistance to l-buthionine-(S,R)-sulfoximine: involvement of Nrf2 activation

Abstract

Pharmacological depletion of l-γ-glutamyl-l-cysteinyl-glycine (GSH) has been implicated in the sensitization of cancer cells to alkylating agents and apoptosis. However, some types of cells do not induce apoptotic response following chemical depletion of GSH. In the present study, we report that murine embryonic fibroblasts (MEFs) can survive in the presence of GSH inhibitor l-buthionine-(S,R)-sulfoximine (BSO), even though most intracellular GSH was depleted. As a cellular adaptive mechanism, BSO treatment effectively activated the NF-E2-related factor 2 (Nrf2) pathway, which led to up-regulation of antioxidant enzymes in these cells through the extracellular signal-regulated kinase cascade. While nrf2-deficient MEFs lost the inducibility of antioxidant genes, which resulted in higher levels of reactive oxygen species accumulation, caspase-3 activation, and cell death than wild-type cells. Finally, nrf2-deficient cells can be more sensitized to doxorubicin-induced cell death by BSO pre-incubation, while wild-type cells were not. In addition, BSO-mediated cell death was facilitated by administering Nrf2 siRNA to chemoresistant human ovarian cancer cells. These results indicate that Nrf2 is the primary factor inducing the cell survival system under GSH depletion and that the effect of BSO as a chemosensitizer might be enhanced by inhibition of Nrf2.

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Abbreviations

GSH:

Glutathione

GCLC:

Catalytic subunit of γ-glutamylcysteine ligase

GCLM:

Modulatory subunit of γ-glutamylcysteine ligase

Nqo1:

NAD(P)H: quinone oxidoreductase 1

ARE:

Antioxidant response element

BSO:

l-Buthionine-(S,R)-sulfoximine

ROS:

Reactive oxygen speices

MAPK:

Mitogen-activated protein kinase

ERK:

Extracellular signal-regulated kinase

JNK:

c-Jun N-terminal kinase

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Acknowledgments

This work was supported by the Korea Science and Engineering Foundation (KOSEF) grant funded by the Korea government (MOST, R01-2007-000-10890-0)

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Correspondence to Mi-Kyoung Kwak.

Additional information

Hyang-Rim Lee and Jeong-Min Cho contributed equally to this work.

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Lee, H., Cho, J., Shin, D. et al. Adaptive response to GSH depletion and resistance to l-buthionine-(S,R)-sulfoximine: involvement of Nrf2 activation. Mol Cell Biochem 318, 23–31 (2008). https://doi.org/10.1007/s11010-008-9853-y

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Keywords

  • GSH
  • BSO
  • Oxidative stress
  • Nrf2
  • Chemosensitizer