Role of Nrf2, HO-1 and GSH in Neuroblastoma Cell Resistance to Bortezomib

A L Furfaro, S. Piras, Cinzia Domenicotti, D Fenoglio, A. De Luigi, M Salmona, L Moretta, Umberto Maria Marinari, Maria Adelaide Pronzato, N. Traverso, Mariapaola Nitti

Research output: Contribution to journalArticle

Abstract

The activation of Nrf2 has been demonstrated to play a crucial role in cancer cell resistance to different anticancer therapies. The inhibition of proteasome activity has been proposed as a chemosensitizing therapy but the activation of Nrf2 could reduce its efficacy. Using the highly chemoresistant neuroblastoma cells HTLA-230, here we show that the strong reduction in proteasome activity, obtained by using low concentration of bortezomib (BTZ, 2.5 nM), fails in reducing cell viability. BTZ treatment favours the binding of Nrf2 to the ARE sequences in the promoter regions of target genes such as heme oxygenase 1 (HO-1), the modulatory subunit of γ-glutamylcysteine ligase (GCLM) and the transporter for cysteine (x-CT), enabling their transcription. GSH level is also increased after BTZ treatment. The up-regulation of Nrf2 target genes is responsible for cell resistance since HO-1 silencing and GSH depletion synergistically decrease BTZ-treated cell viability. Moreover, cell exposure to all-trans-Retinoic acid (ATRA, 3 μM) reduces the binding of Nrf2 to the ARE sequences, decreases HO-1 induction and lowers GSH level increasing the efficacy of bortezomib. These data suggest the role of Nrf2, HO-1 and GSH as molecular targets to improve the efficacy of low doses of bortezomib in the treatment of malignant neuroblastoma.

Original languageEnglish
Pages (from-to)e0152465
JournalPLoS One
Volume11
Issue number3
DOIs
Publication statusPublished - 2016

Fingerprint

heme oxygenase (biliverdin-producing)
Heme Oxygenase-1
Neuroblastoma
Cells
proteasome endopeptidase complex
Proteasome Endopeptidase Complex
cell viability
Cell Survival
Genes
Chemical activation
cells
therapeutics
retinoic acid
Transcription
Ligases
Tretinoin
ligases
Genetic Promoter Regions
Cysteine
cysteine

Keywords

  • Amino Acid Transport System y+
  • Antioxidant Response Elements
  • Bortezomib
  • Cell Line, Tumor
  • Cell Survival
  • Dose-Response Relationship, Drug
  • Drug Resistance, Neoplasm
  • Gene Expression Regulation, Neoplastic
  • Gene Silencing
  • Glutamate-Cysteine Ligase
  • Glutathione
  • Heme Oxygenase-1
  • Humans
  • NF-E2-Related Factor 2
  • Neuroblastoma
  • Proteasome Endopeptidase Complex
  • Protein Subunits
  • RNA, Messenger
  • Transcription, Genetic
  • Tretinoin
  • Up-Regulation
  • Journal Article
  • Research Support, Non-U.S. Gov't

Cite this

Role of Nrf2, HO-1 and GSH in Neuroblastoma Cell Resistance to Bortezomib. / Furfaro, A L; Piras, S.; Domenicotti, Cinzia; Fenoglio, D; De Luigi, A.; Salmona, M; Moretta, L; Marinari, Umberto Maria; Pronzato, Maria Adelaide; Traverso, N.; Nitti, Mariapaola.

In: PLoS One, Vol. 11, No. 3, 2016, p. e0152465.

Research output: Contribution to journalArticle

Furfaro, AL, Piras, S, Domenicotti, C, Fenoglio, D, De Luigi, A, Salmona, M, Moretta, L, Marinari, UM, Pronzato, MA, Traverso, N & Nitti, M 2016, 'Role of Nrf2, HO-1 and GSH in Neuroblastoma Cell Resistance to Bortezomib', PLoS One, vol. 11, no. 3, pp. e0152465. https://doi.org/10.1371/journal.pone.0152465
Furfaro, A L ; Piras, S. ; Domenicotti, Cinzia ; Fenoglio, D ; De Luigi, A. ; Salmona, M ; Moretta, L ; Marinari, Umberto Maria ; Pronzato, Maria Adelaide ; Traverso, N. ; Nitti, Mariapaola. / Role of Nrf2, HO-1 and GSH in Neuroblastoma Cell Resistance to Bortezomib. In: PLoS One. 2016 ; Vol. 11, No. 3. pp. e0152465.
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AU - Furfaro, A L

AU - Piras, S.

AU - Domenicotti, Cinzia

AU - Fenoglio, D

AU - De Luigi, A.

AU - Salmona, M

AU - Moretta, L

AU - Marinari, Umberto Maria

AU - Pronzato, Maria Adelaide

AU - Traverso, N.

AU - Nitti, Mariapaola

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N2 - The activation of Nrf2 has been demonstrated to play a crucial role in cancer cell resistance to different anticancer therapies. The inhibition of proteasome activity has been proposed as a chemosensitizing therapy but the activation of Nrf2 could reduce its efficacy. Using the highly chemoresistant neuroblastoma cells HTLA-230, here we show that the strong reduction in proteasome activity, obtained by using low concentration of bortezomib (BTZ, 2.5 nM), fails in reducing cell viability. BTZ treatment favours the binding of Nrf2 to the ARE sequences in the promoter regions of target genes such as heme oxygenase 1 (HO-1), the modulatory subunit of γ-glutamylcysteine ligase (GCLM) and the transporter for cysteine (x-CT), enabling their transcription. GSH level is also increased after BTZ treatment. The up-regulation of Nrf2 target genes is responsible for cell resistance since HO-1 silencing and GSH depletion synergistically decrease BTZ-treated cell viability. Moreover, cell exposure to all-trans-Retinoic acid (ATRA, 3 μM) reduces the binding of Nrf2 to the ARE sequences, decreases HO-1 induction and lowers GSH level increasing the efficacy of bortezomib. These data suggest the role of Nrf2, HO-1 and GSH as molecular targets to improve the efficacy of low doses of bortezomib in the treatment of malignant neuroblastoma.

AB - The activation of Nrf2 has been demonstrated to play a crucial role in cancer cell resistance to different anticancer therapies. The inhibition of proteasome activity has been proposed as a chemosensitizing therapy but the activation of Nrf2 could reduce its efficacy. Using the highly chemoresistant neuroblastoma cells HTLA-230, here we show that the strong reduction in proteasome activity, obtained by using low concentration of bortezomib (BTZ, 2.5 nM), fails in reducing cell viability. BTZ treatment favours the binding of Nrf2 to the ARE sequences in the promoter regions of target genes such as heme oxygenase 1 (HO-1), the modulatory subunit of γ-glutamylcysteine ligase (GCLM) and the transporter for cysteine (x-CT), enabling their transcription. GSH level is also increased after BTZ treatment. The up-regulation of Nrf2 target genes is responsible for cell resistance since HO-1 silencing and GSH depletion synergistically decrease BTZ-treated cell viability. Moreover, cell exposure to all-trans-Retinoic acid (ATRA, 3 μM) reduces the binding of Nrf2 to the ARE sequences, decreases HO-1 induction and lowers GSH level increasing the efficacy of bortezomib. These data suggest the role of Nrf2, HO-1 and GSH as molecular targets to improve the efficacy of low doses of bortezomib in the treatment of malignant neuroblastoma.

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KW - Humans

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KW - Proteasome Endopeptidase Complex

KW - Protein Subunits

KW - RNA, Messenger

KW - Transcription, Genetic

KW - Tretinoin

KW - Up-Regulation

KW - Journal Article

KW - Research Support, Non-U.S. Gov't

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DO - 10.1371/journal.pone.0152465

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SP - e0152465

JO - PLoS One

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SN - 1932-6203

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