Aberrant regulation of the MRP3 gene in non-small cell lung carcinoma

Christopher M. Mahaffey, Nichole C. Mahaffey, William Holland, Hongqiao Zhang, David R Gandara, Philip Mack, Henry Jay Forman

Research output: Contribution to journalArticle

11 Citations (Scopus)

Abstract

Introduction: Multidrug-resistant protein-3 (MRP3), a membrane-bound transporter, facilitates efflux of toxic compounds, including certain chemotherapies, out of cells. Aberrant MRP3 expression has been linked to drug resistance in non-small cell lung carcinoma (NSCLC). We sought to determine if tumor MRP3 expression patterns correlate with the mutational status of upstream regulators, including nuclear factor erythroid-2-related factor 2 (Nrf2) and its functional repressor Keap1 in NSCLC cell lines and patient samples. Methods: To identify putative Nrf2-binding sites in the MRP3 promoter and to evaluate Keap1, Nrf2, and p53 mutation status in four cell lines and 33 NSCLC surgically resected tumor specimens with regard to their impact on MRP3 levels. Results: Chromatin immunoprecipitation analysis of the MRP3 promoter revealed an almost threefold increase in Nrf2 binding to the third putative Nrf2-binding sequence distal to the start site, demonstrating direct regulation of MRP3 by Nrf2. In NSCLC cell lines, elevated Nrf2 protein was observed in cell lines with increased MRP3 RNA expression. In patient tumor specimens, the presence of mutations in Keap1/Nrf2 correlated with MRP3 RNA levels (p < 0.05). p53 mutations were observed in 33% of cases, and all Keap1 mutant-positive tumors possessed a p53 mutation (n = 5; p = 0.0019). Conclusions: We demonstrate direct involvement between the transcription factor Nrf2 and the MRP3 promoter, which leads to the up-regulation of the MRP3 gene. In addition, we found a statistically significant correlation between the presence of Keap1/Nrf2 mutations and increased MRP3 messenger RNA levels in our NSCLC patient samples.

Original languageEnglish (US)
Pages (from-to)34-39
Number of pages6
JournalJournal of Thoracic Oncology
Volume7
Issue number1
DOIs
StatePublished - Jan 2012

Fingerprint

Non-Small Cell Lung Carcinoma
Proteins
Mutation
Cell Line
Neoplasms
RNA
Membrane Transport Proteins
Chromatin Immunoprecipitation
Poisons
Drug Resistance
Transcription Factors
Up-Regulation
Binding Sites
Drug Therapy

Keywords

  • Keap1
  • MRP3
  • Nrf2
  • NSCLC
  • p53

ASJC Scopus subject areas

  • Oncology
  • Pulmonary and Respiratory Medicine

Cite this

Aberrant regulation of the MRP3 gene in non-small cell lung carcinoma. / Mahaffey, Christopher M.; Mahaffey, Nichole C.; Holland, William; Zhang, Hongqiao; Gandara, David R; Mack, Philip; Forman, Henry Jay.

In: Journal of Thoracic Oncology, Vol. 7, No. 1, 01.2012, p. 34-39.

Research output: Contribution to journalArticle

Mahaffey, Christopher M. ; Mahaffey, Nichole C. ; Holland, William ; Zhang, Hongqiao ; Gandara, David R ; Mack, Philip ; Forman, Henry Jay. / Aberrant regulation of the MRP3 gene in non-small cell lung carcinoma. In: Journal of Thoracic Oncology. 2012 ; Vol. 7, No. 1. pp. 34-39.
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abstract = "Introduction: Multidrug-resistant protein-3 (MRP3), a membrane-bound transporter, facilitates efflux of toxic compounds, including certain chemotherapies, out of cells. Aberrant MRP3 expression has been linked to drug resistance in non-small cell lung carcinoma (NSCLC). We sought to determine if tumor MRP3 expression patterns correlate with the mutational status of upstream regulators, including nuclear factor erythroid-2-related factor 2 (Nrf2) and its functional repressor Keap1 in NSCLC cell lines and patient samples. Methods: To identify putative Nrf2-binding sites in the MRP3 promoter and to evaluate Keap1, Nrf2, and p53 mutation status in four cell lines and 33 NSCLC surgically resected tumor specimens with regard to their impact on MRP3 levels. Results: Chromatin immunoprecipitation analysis of the MRP3 promoter revealed an almost threefold increase in Nrf2 binding to the third putative Nrf2-binding sequence distal to the start site, demonstrating direct regulation of MRP3 by Nrf2. In NSCLC cell lines, elevated Nrf2 protein was observed in cell lines with increased MRP3 RNA expression. In patient tumor specimens, the presence of mutations in Keap1/Nrf2 correlated with MRP3 RNA levels (p < 0.05). p53 mutations were observed in 33{\%} of cases, and all Keap1 mutant-positive tumors possessed a p53 mutation (n = 5; p = 0.0019). Conclusions: We demonstrate direct involvement between the transcription factor Nrf2 and the MRP3 promoter, which leads to the up-regulation of the MRP3 gene. In addition, we found a statistically significant correlation between the presence of Keap1/Nrf2 mutations and increased MRP3 messenger RNA levels in our NSCLC patient samples.",
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