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BRCA1 modulates sensitivity to 5F-203 by regulating xenobiotic stress-inducible protein levels and EROD activity

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Abstract

Purpose

We have investigated the effects of BRCA1 over-expression and knockdown on 5F-203-induced gene expression and cytotoxicity in human breast cancer cells. 5F-203 is a chemotherapeutic prodrug that both induces a p450 enzyme, CYP1A1, and is metabolically activated by CYP1A1.

Methods

We used several molecular biological techniques to confirm our findings. BRCA1 regulates sensitivity to 5F-203 by regulating the expression of CYP1A1 mRNA and its EROD activity. XRE-Luc reporter assays, semi-quantitative RT-PCR, Western blot analysis, EROD activity measurements, gene knockdown and MTT cell survival assays were used for this study.

Results

Our results show that the ability of 5F-203 treatments to increase CYP1A1 mRNA level and CYP1A1 enzymatic activity (EROD activity) are affected by BRCA1 protein levels. In addition, the ability of 5F-203 treatments to induce proteins, P53 and P53 target genes such as P21, is significantly decreased in BRCA1 knockdown cells, suggesting that BRCA1-related effects could at least partially explain why BRCA1 knockdown increases resistance to 5F-203-mediated cytotoxicity. We also observed altered expression of the two major transcription factors (AhR and ARNT) that affect CYP1A1 expression when BRCA1 protein levels are altered.

Conclusion

BRCA1 is an important protein, which affects 5F-203-mediated cytotoxicity. Our findings are potentially clinically significant; they suggest that those patients most likely to respond to this new prodrug will have tumors containing normal amounts of BRCA1.

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Abbreviations

AhR:

Aryl hydrocarbon receptor

BRCA1:

Breast cancer susceptibility gene-1

DMEM:

Dulbecco’s modified Eagles’ medium

DMSO:

Dimethyl sulfoxide

FBS:

Fetal bovine serum

SEM:

Standard error of mean

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Acknowledgments

Dr. Bae was supported, in part, by the National Institute of Environmental Health Science, NIH (ES01440-01), US Department of Defense (DOD) Breast Cancer Program Idea Award (DAMD17-02-1-0525), American Cancer Society (IRG 97-152-13), and Susan G. Komen for the Cure (BCTR119906 and FAS0703858). We appreciate Dr. Thomas L. Mattson for his helpful discussions. Dr. Lee was partially supported by Soonchungyang University during his sabbatical year (2007) in Georgetown University.

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Authors and Affiliations

  1. Department of Oncology, Lombardi Comprehensive Cancer Center, Georgetown University, 3970 Reservoir Road, NW, Washington DC, 20057-1469, USA

    Hyo Jin Kang, Hee Jeong Kim, Sang Hoon Kwon, Sang Han Lee & Insoo Bae

  2. Department of Radiation Medicine, Lombardi Comprehensive Cancer Center, Georgetown University, 3970 Reservoir Road, NW, Washington DC, USA

    Insoo Bae

  3. Department of Obstetrics and Gynecology, Keimyung University, School of Medicine, Daegu, Korea

    Sang Hoon Kwon

  4. Department of Biochemistry, University of Michigan, Ann Arbor, MI, USA

    Brian DongHoon Kang

  5. Molecular Carcinogenesis, National Institute of Environmental Health Sciences, National Institute of Health, Research Triangle Park, NC, USA

    Thomas E. Eling

  6. Department of Biochemistry, College of Medicine, Soonchunhyang University, Cheon-An, Korea

    Sang Han Lee

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  1. Hyo Jin Kang
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  2. Hee Jeong Kim
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  3. Sang Hoon Kwon
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  4. Brian DongHoon Kang
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  5. Thomas E. Eling
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  6. Sang Han Lee
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  7. Insoo Bae
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Correspondence to Insoo Bae.

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Kang, H.J., Kim, H.J., Kwon, S.H. et al. BRCA1 modulates sensitivity to 5F-203 by regulating xenobiotic stress-inducible protein levels and EROD activity . Cancer Chemother Pharmacol 62, 689–697 (2008). https://doi.org/10.1007/s00280-007-0657-7

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  • DOI: https://doi.org/10.1007/s00280-007-0657-7

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