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Pharmacogenomics of Phenolic Antioxidant Butylated Hydroxyanisole (BHA) in the Small Intestine and Liver of Nrf2 Knockout and C57BL/6J Mice

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Abstract

Purpose

The objective of this study was to investigate the pharmacogenomics and the spatial regulation of global gene expression profiles elicited by cancer chemopreventive agent butylated hydroxyanisole (BHA) in mouse small intestine and liver as well as to identify BHA-modulated nuclear factor-E2-related factor 2 (Nrf2)-dependent genes.

Methods

C57BL/6J (+/+; wildtype) and C57BL/6J/Nrf2(−/−; knockout) mice were administered a single 200 mg/kg oral dose of BHA or only vehicle. Both small intestine and liver were collected at 3 h after treatment and total RNA was extracted. Gene expression profiles were analyzed using 45,000 Affymetrix mouse genome 430 2.0 array and GeneSpring 7.2 software. Microarray results were validated by quantitative real-time reverse transcription-PCR analyses.

Results

Clusters of genes that were either induced or suppressed more than two fold by BHA treatment compared with vehicle in C57BL/6J/Nrf2(−/−; knockout) and C57BL/6J Nrf2 (+/+; wildtype) mice genotypes were identified. Amongst these, in small intestine and liver, 1,490 and 493 genes respectively were identified as Nrf2-dependent and upregulated, and 1,090 and 824 genes respectively as Nrf2-dependent and downregulated. Based on their biological functions, these genes can be categorized into ubiquitination/proteolysis, apoptosis/cell cycle, electron transport, detoxification, cell growth/differentiation, transcription factors/interacting partners, kinases and phosphatases, transport, biosynthesis/metabolism, RNA/protein processing and nuclear assembly, and DNA replication genes. Phase II detoxification/antioxidant genes as well as novel molecular target genes, including putative interacting partners of Nrf2 such as nuclear corepressors and coactivators, were also identified as Nrf2-dependent genes.

Conclusions

The identification of BHA-regulated and Nrf2-dependent genes not only provides potential novel insights into the gestalt biological effects of BHA on the pharmacogenomics and spatial regulation of global gene expression profiles in cancer chemoprevention, but also points to the pivotal role of Nrf2 in these biological processes.

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Abbreviations

ARE:

antioxidant response element

BHA:

butylated hydroxyanisole

Mapk:

mitogen-activated protein kinase

Nrf2:

nuclear factor-E2 -related factor 2

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Acknowledgments

The authors are deeply grateful to Mr. Curtis Krier at the Cancer Institute of New Jersey (CINJ) Core Expression Array Facility for his expert assistance with the microarray analyses. The authors are also deeply indebted to Ms. Donna Wilson of the Keck Center for Collaborative Neuroscience, Rutgers University as well as the staff of the Human Genetics Institute of New Jersey at Rutgers University for their great expertise and help with the quantitative real-time PCR analyses. This work was supported in part by NIH grant R01-CA094828.

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Authors and Affiliations

  1. Graduate Program in Pharmaceutical Sciences, Department of Pharmaceutics, Ernest Mario School of Pharmacy, Rutgers, The State University of New Jersey, Piscataway, New Jersey, 08854, USA

    Sujit Nair, Guoxiang Shen, Avantika Gopalakrishnan,  Rong Hu, Wen Lin & Young-Sam Keum

  2. Department of Pharmaceutics, Ernest Mario School of Pharmacy, Rutgers, The State University of New Jersey, Piscataway, New Jersey, 08854, USA

    Changjiang Xu, Vidya Hebbar & Ah-Ng Tony Kong

  3. Department of Biochemistry and Molecular Biology, UMDNJ-New Jersey Medical School, 185 South Orange Avenue, Newark, New Jersey, 07103, USA

    Mohit Raja Jain

  4. Department of Pharmacy, National University of Singapore, 18, Science Drive 4, Singapore, 117543, Singapore

    Celine Liew

  5. Department of Pathology, University of California, D440 Medical Sciences, Irvine, California, 92697, USA

    Jefferson Y. Chan

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  1. Sujit Nair
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  2. Changjiang Xu
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  12. Ah-Ng Tony Kong
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Correspondence to Ah-Ng Tony Kong.

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Sujit Nair and Changjiang Xu contributed equally to the present study.

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Nair, S., Xu, C., Shen, G. et al. Pharmacogenomics of Phenolic Antioxidant Butylated Hydroxyanisole (BHA) in the Small Intestine and Liver of Nrf2 Knockout and C57BL/6J Mice. Pharm Res 23, 2621–2637 (2006). https://doi.org/10.1007/s11095-006-9099-x

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  • DOI: https://doi.org/10.1007/s11095-006-9099-x

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