Multidrug-resistant protein-3 gene regulation by the transcription factor Nrf2 in human bronchial epithelial and non-small-cell lung carcinoma

Christopher M. Mahaffey, Hongqiao Zhang, Alessandra Rinna, William Holland, Philip Mack, Henry Jay Forman

Research output: Contribution to journalArticle

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Abstract

Multidrug-resistant proteins (MRPs) are members of the ATP-binding cassette superfamily that facilitate detoxification by transporting toxic compounds, including chemotherapeutic drugs, out of cells. Chemotherapy, radiation, and other xenobiotic stresses have been shown to increase levels of select MRPs, although the underlying mechanism remains largely unknown. Additionally, MRP3 is suspected of playing a role in the drug resistance of non-small-cell lung carcinoma (NSCLC). Analysis of the MRP3 promoter revealed the presence of multiple putative electrophile-responsive elements (EpREs), sequences that suggest possible regulation of this gene by Nrf2, the key transcription factor that binds to EpRE. The goal of this investigation was to determine whether MRP3 induction was dependent upon the transcription factor Nrf2. Keap1, a key regulator of Nrf2, sequesters Nrf2 in the cytoplasm, preventing entry into the nucleus. The electrophilic lipid peroxidation product 4-hydroxy-2-nonenal (HNE) has been shown to modify Keap1, allowing Nrf2 to enter the nucleus. We found that HNE up-regulated MRP3 mRNA and protein levels in cell lines with wild-type Keap1 (the human bronchial epithelial cell line HBE1 and the NSCLC cell line H358), but not in the Keap1-mutant NSCLC cell lines (A549 and H460). Cell lines with mutant Keap1 had constitutively higher MRP3 that was not increased by HNE treatment. In HBE1 cells, silencing of Nrf2 with siRNA inhibited induction of MRP3 by HNE. Finally, we found that silencing Nrf2 also increased the toxicity of cisplatin in H358 cells. The combined results therefore support the hypothesis that MRP3 induction by HNE involves Nrf2 activation.

Original languageEnglish (US)
Pages (from-to)1650-1657
Number of pages8
JournalFree Radical Biology and Medicine
Volume46
Issue number12
DOIs
StatePublished - Jun 15 2009

Fingerprint

Gene expression
Non-Small Cell Lung Carcinoma
Transcription Factors
Cells
Cell Line
Proteins
Detoxification
Chemotherapy
Poisons
Xenobiotics
Drug Resistance
Pharmaceutical Preparations
Small Interfering RNA
Cisplatin
Lipid Peroxidation
Toxicity
Cytoplasm
Genes
Adenosine Triphosphate
Epithelial Cells

Keywords

  • 4-Hydroxynonenal
  • A549 cell line
  • Electrophile response element
  • Free radicals
  • H358 cell line
  • H460 cell line
  • HBE1 cell line
  • Keap1
  • Lung cancer
  • multidrug-resistant proteins
  • Nrf2

ASJC Scopus subject areas

  • Biochemistry
  • Physiology (medical)

Cite this

Multidrug-resistant protein-3 gene regulation by the transcription factor Nrf2 in human bronchial epithelial and non-small-cell lung carcinoma. / Mahaffey, Christopher M.; Zhang, Hongqiao; Rinna, Alessandra; Holland, William; Mack, Philip; Forman, Henry Jay.

In: Free Radical Biology and Medicine, Vol. 46, No. 12, 15.06.2009, p. 1650-1657.

Research output: Contribution to journalArticle

Mahaffey, Christopher M. ; Zhang, Hongqiao ; Rinna, Alessandra ; Holland, William ; Mack, Philip ; Forman, Henry Jay. / Multidrug-resistant protein-3 gene regulation by the transcription factor Nrf2 in human bronchial epithelial and non-small-cell lung carcinoma. In: Free Radical Biology and Medicine. 2009 ; Vol. 46, No. 12. pp. 1650-1657.
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