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Aberrant Regulation of the MRP3 Gene in Non-small Cell Lung Carcinoma

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Presentation on theme: "Aberrant Regulation of the MRP3 Gene in Non-small Cell Lung Carcinoma"— Presentation transcript:

1 Aberrant Regulation of the MRP3 Gene in Non-small Cell Lung Carcinoma
Christopher M. Mahaffey, PhD, Nichole C. Mahaffey, PhD, William Holland, MS, Hongqiao Zhang, PhD, David R. Gandara, Philip C. Mack, PhD, Henry Jay Forman, PhD  Journal of Thoracic Oncology  Volume 7, Issue 1, Pages (January 2012) DOI: /JTO.0b013e318233d753 Copyright © 2012 International Association for the Study of Lung Cancer Terms and Conditions

2 FIGURE 1 EpRE-binding sites are located −434, −628, −805, and −1049 bp. Nrf2 binding was increased ∼2.8-fold versus control at the −805 bp EpRE after 3 hours exposure to 15μM HNE. EpRE, electrophilic response element; Nrf2, nuclear factor erythroid-2-related factor 2; HNE, 4-hydroxy-2-nonenal. Journal of Thoracic Oncology 2012 7, 34-39DOI: ( /JTO.0b013e318233d753) Copyright © 2012 International Association for the Study of Lung Cancer Terms and Conditions

3 FIGURE 2 (A) PCR amplification of p53. Exons 5 to 8 were successfully amplified in patient samples. (B) Single-strand conformation polymorphism (SSCP). Mutations were found in 11 of 33 samples and confirmed by sequencing. (C) The left column represents the number of p53 wt patient samples in our cohort (22/33), and the right column is the number of p53 mt patient samples (11/33). All five Keap1 mutations occurred in conjunction with a p53 mutation; Fisher's exact test = two-sided p value is PCR, polymerase chain reaction; mt, mutant. Journal of Thoracic Oncology 2012 7, 34-39DOI: ( /JTO.0b013e318233d753) Copyright © 2012 International Association for the Study of Lung Cancer Terms and Conditions

4 FIGURE 3 (A) H1650 (p53 wt/Keap1 wt/Nrf2 wt), H358 (p53 null/Keap1 wt/Nrf2 wt), H1666 (p53 wt/Keap1 wt/Nrf2 wt), A549 (p53 wt/Keap1 mt/Nrf2 wt), and DU-145 (p53 mt/Keap1 wt*). Levels of MRP3 mRNA of our cell line model are compared. (B) Western blot of standard and ∼110 KD Nrf2 in our cell line model. (C) Scion image absorbance measurements. Levels of total ∼67 KD and ∼110 KD Nrf2 as a ratio to respective β-actin levels. Horizontal lines = Keap1 wt, black = Keap1 mt, gray shading = Keap1 splicing errors. Results are expressed as means ± SEM (n = 3). MRP3, mRNA, messenger RNA; Nrf2, nuclear factor erythroid-2-related factor 2; SEM, standard error of the mean. Journal of Thoracic Oncology 2012 7, 34-39DOI: ( /JTO.0b013e318233d753) Copyright © 2012 International Association for the Study of Lung Cancer Terms and Conditions

5 FIGURE 4 Patient MRP3 mRNA levels were normalized to those of H358. Keap1/Nrf2 wt patient samples (Mean ∼2.6-fold versus H358) and Keap1/Nrf2 homozygous mt patient samples (Mean ∼5.6-fold versus H358) were significantly different (p < 0.05). Results are expressed as means ± SEM. MRP3, multidrug-resistant protein-3; mRNA, messenger RNA; mt, mutant; SEM, standard error of the mean. Journal of Thoracic Oncology 2012 7, 34-39DOI: ( /JTO.0b013e318233d753) Copyright © 2012 International Association for the Study of Lung Cancer Terms and Conditions

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