Overexpression of NQO1 protects human SK-N-MC neuroblastoma cells against dopamine-induced cell death

K. S. Zafar, S. H. Inayat-Hussain, D. Siegel, A. Bao, B. Shieh, D. Ross

Research output: Contribution to journalArticle

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Abstract

NAD(P)H quinone oxidoreductase 1 (NQO1) can metabolize dopamine-derived quinones (DAQ) and absence of NQO1 due to the NQO1*2 polymorphism has been suggested to be a risk factor for Parkinson's disease. In order to define whether NQO1 plays a protective role in dopamine toxicity, we have examined the potential role of NQO1 in the SK-N-MC human neuroblastoma cell line. SK-N-MC cells were stably transfected with NQO1 to generate stable clones with NQO1 enzymatic activity of 245 nmol/mg min while vector control and parental cells had NQO1 activities of less than 12 nmol/mg min. Incubation of dopamine for 24 h in both parental and vector control SK-N-MC cells resulted in 85% and 72% cell death as assessed by annexin-V/propidium iodide analysis. In agreement, 88% and 84% of parental and vector control cells, respectively underwent loss of mitochondrial membrane potential (MMP) assessed by tetramethylrhodamine ethyl ester. In contrast, NQO1-transfected cells were resistant to dopamine toxicity and both cell death and loss of MMP were markedly abrogated in NQO1-transfected SK-N-MC cells. When dopamine was added to medium, oxygen uptake could be detected indicating autoxidation with concomitant formation of oxygen radicals and quinones. However, dopamine-induced cell death was not affected by the inclusion of either superoxide dismutase or catalase suggesting that superoxide and hydrogen peroxide were not involved in toxicity. Quinones formed in medium may exert toxicity extracellularly or intracellularly but the protective role of NQO1 argues for an intracellular mechanism. In summary, transfection of SK-N-MC cells with NQO1 protects against dopamine-induced toxicity.

Original languageEnglish
Pages (from-to)261-267
Number of pages7
JournalToxicology Letters
Volume166
Issue number3
DOIs
Publication statusPublished - 25 Oct 2006
Externally publishedYes

Fingerprint

Cell death
Neuroblastoma
Dopamine
Cell Death
Toxicity
Quinones
Mitochondrial Membrane Potential
Membranes
Propidium
Annexin A5
Polymorphism
Superoxides
NAD
Catalase
Hydrogen Peroxide
Superoxide Dismutase
Transfection
Parkinson Disease
Reactive Oxygen Species
Oxidoreductases

Keywords

  • Apoptosis
  • Dopamine
  • DT-diaphorase
  • NQO1
  • Parkinson's disease
  • Quinones

ASJC Scopus subject areas

  • Toxicology

Cite this

Zafar, K. S., Inayat-Hussain, S. H., Siegel, D., Bao, A., Shieh, B., & Ross, D. (2006). Overexpression of NQO1 protects human SK-N-MC neuroblastoma cells against dopamine-induced cell death. Toxicology Letters, 166(3), 261-267. https://doi.org/10.1016/j.toxlet.2006.07.340

Overexpression of NQO1 protects human SK-N-MC neuroblastoma cells against dopamine-induced cell death. / Zafar, K. S.; Inayat-Hussain, S. H.; Siegel, D.; Bao, A.; Shieh, B.; Ross, D.

In: Toxicology Letters, Vol. 166, No. 3, 25.10.2006, p. 261-267.

Research output: Contribution to journalArticle

Zafar, KS, Inayat-Hussain, SH, Siegel, D, Bao, A, Shieh, B & Ross, D 2006, 'Overexpression of NQO1 protects human SK-N-MC neuroblastoma cells against dopamine-induced cell death', Toxicology Letters, vol. 166, no. 3, pp. 261-267. https://doi.org/10.1016/j.toxlet.2006.07.340
Zafar, K. S. ; Inayat-Hussain, S. H. ; Siegel, D. ; Bao, A. ; Shieh, B. ; Ross, D. / Overexpression of NQO1 protects human SK-N-MC neuroblastoma cells against dopamine-induced cell death. In: Toxicology Letters. 2006 ; Vol. 166, No. 3. pp. 261-267.
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abstract = "NAD(P)H quinone oxidoreductase 1 (NQO1) can metabolize dopamine-derived quinones (DAQ) and absence of NQO1 due to the NQO1*2 polymorphism has been suggested to be a risk factor for Parkinson's disease. In order to define whether NQO1 plays a protective role in dopamine toxicity, we have examined the potential role of NQO1 in the SK-N-MC human neuroblastoma cell line. SK-N-MC cells were stably transfected with NQO1 to generate stable clones with NQO1 enzymatic activity of 245 nmol/mg min while vector control and parental cells had NQO1 activities of less than 12 nmol/mg min. Incubation of dopamine for 24 h in both parental and vector control SK-N-MC cells resulted in 85{\%} and 72{\%} cell death as assessed by annexin-V/propidium iodide analysis. In agreement, 88{\%} and 84{\%} of parental and vector control cells, respectively underwent loss of mitochondrial membrane potential (MMP) assessed by tetramethylrhodamine ethyl ester. In contrast, NQO1-transfected cells were resistant to dopamine toxicity and both cell death and loss of MMP were markedly abrogated in NQO1-transfected SK-N-MC cells. When dopamine was added to medium, oxygen uptake could be detected indicating autoxidation with concomitant formation of oxygen radicals and quinones. However, dopamine-induced cell death was not affected by the inclusion of either superoxide dismutase or catalase suggesting that superoxide and hydrogen peroxide were not involved in toxicity. Quinones formed in medium may exert toxicity extracellularly or intracellularly but the protective role of NQO1 argues for an intracellular mechanism. In summary, transfection of SK-N-MC cells with NQO1 protects against dopamine-induced toxicity.",
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AU - Bao, A.

AU - Shieh, B.

AU - Ross, D.

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AB - NAD(P)H quinone oxidoreductase 1 (NQO1) can metabolize dopamine-derived quinones (DAQ) and absence of NQO1 due to the NQO1*2 polymorphism has been suggested to be a risk factor for Parkinson's disease. In order to define whether NQO1 plays a protective role in dopamine toxicity, we have examined the potential role of NQO1 in the SK-N-MC human neuroblastoma cell line. SK-N-MC cells were stably transfected with NQO1 to generate stable clones with NQO1 enzymatic activity of 245 nmol/mg min while vector control and parental cells had NQO1 activities of less than 12 nmol/mg min. Incubation of dopamine for 24 h in both parental and vector control SK-N-MC cells resulted in 85% and 72% cell death as assessed by annexin-V/propidium iodide analysis. In agreement, 88% and 84% of parental and vector control cells, respectively underwent loss of mitochondrial membrane potential (MMP) assessed by tetramethylrhodamine ethyl ester. In contrast, NQO1-transfected cells were resistant to dopamine toxicity and both cell death and loss of MMP were markedly abrogated in NQO1-transfected SK-N-MC cells. When dopamine was added to medium, oxygen uptake could be detected indicating autoxidation with concomitant formation of oxygen radicals and quinones. However, dopamine-induced cell death was not affected by the inclusion of either superoxide dismutase or catalase suggesting that superoxide and hydrogen peroxide were not involved in toxicity. Quinones formed in medium may exert toxicity extracellularly or intracellularly but the protective role of NQO1 argues for an intracellular mechanism. In summary, transfection of SK-N-MC cells with NQO1 protects against dopamine-induced toxicity.

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